Imidazolopyrazine compounds useful for the treatment of degenerative and inflammatory diseases

ABSTRACT

Novel imidazo[ 1,2 -a]pyrazine compounds are disclosed that have a formula represented by the following: 
                         
The compounds may be prepared as pharmaceutical compositions, and may be used for the prevention and treatment of a variety of conditions in mammals including humans, including by way of non-limiting example, arthritis, inflammation, and others.

CROSS REFERENCE TO RELATED APPLICATIONS

The present application claims the priority of co-pending provisionalapplication U.S. Ser. No. 60/747,223 filed on May 15, 2006, and Ser.Nos. 60/928,539, 60/928,600 60/928,639 and 60/928,568, all bearing thesame title as the present application, and filed on May 10, 2007, andthe disclosures of such applications are incorporated by referenceherein in their entireties. Applicants claim the benefits of suchapplications under 35 U.S.C. §119(e).

FIELD OF THE INVENTION

The present invention relates to a class of imidazolopyrazine compoundscapable of binding to the active site of a serine/threonine kinase, theexpression of which is involved in the pathway resulting in thedegradation of extra-cellular matrix (ECM), joint degeneration anddiseases involving such degradation and/or inflammation.

Diseases involving the degradation of extra-cellular matrix include, butare not limited to, psoriatic arthritis, juvenile arthritis, earlyarthritis, reactive arthritis, osteoarthritis, ankylosing spondylitis,osteoporosis, muskulo skeletal diseases like tendonitis and periodontaldisease, cancer metastasis, airway diseases (COPD, asthma), renal andliver fibrosis, cardio-vascular diseases like atherosclerosis and heartfailure, and neurological diseases like neuroinflammation and multiplesclerosis. Diseases involving primarily joint degeneration include, butare not limited to, psoriatic arthritis, juvenile arthritis, earlyarthritis, reactive arthritis, osteoarthritis, and ankylosingspondylitis.

Rheumatoid arthritis (RA) is a chronic joint degenerative disease,characterized by inflammation and destruction of the joint structures.When the disease is unchecked, it leads to substantial disability andpain due to loss of joint functionality and even premature death. Theaim of an RA therapy, therefore, is not to slow down the disease but toattain remission in order to stop the joint destruction. Besides theseverity of the disease outcome, the high prevalence of RA (˜0.8% of theadults are affected worldwide) means a high socio-economic impact. (Forreviews on RA, we refer to Smolen and Steiner (2003); Lee and Weinblatt(2001); Choy and Panayi (2001); O'Dell (2004) and Firestein (2003)).

Although it is widely accepted that RA is an auto-immune disease, thereis no consensus concerning the precise mechanisms driving the‘initiation stage’ of the disease. What is known is that the initialtrigger(s) does mediate, in a predisposed host, a cascade of events thatleads to the activation of various cell types (B-cells, T-cells,macrophages, fibroblasts, endothelial cells, dendritic cells andothers). Concomitantly, an increased production of various cytokines isobserved in the joints and tissues surrounding the joint (e.g. TNF-α,IL-6, IL-1, IL-15, IL-18 and others). When the disease progresses, thecellular activation and cytokine production cascade becomesself-perpetuating. At this early stage, the destruction of jointstructures is already very clear at this early stage. Thirty percent ofthe patients have radiographic evidence of bony erosions at the time ofdiagnosis and this proportion increases to 60 percent after two years.

Histological analysis of the joints of RA patients clearly evidences themechanisms involved in the RA-associated degradative processes. Thisanalysis shows that the main effector responsible for RA-associatedjoint degradation is the pannus, where the synovial fibroblast, byproducing diverse proteolytic enzymes, is the prime driver of cartilageand bone erosion. A joint classically contains two adjacent bones thatarticulate on a cartilage layer and are surrounded by the synovialmembrane and joint capsule. In the advanced RA patient, the synovium ofjoint increases in size to form the pannus, due to the proliferation ofthe synovial fibroblasts and the infiltration of mononuclear cells suchas T-cells, B-cells, monocytes, macrophages and neutrophils. The pannusmediates the degradation of the adjacent cartilage, leading to thenarrowing of the joint space, and has the potential to invade adjacentbone and cartilage. As bone and cartilage tissues are composed mainly ofcollagen type I or II, respectively, the pannus destructive and invasiveproperties are mediated by the secretion of collagenolytic proteases,principally the matrix metallo proteinases (MMPs). The erosion of thebone under and adjacent to the cartilage is also part of the RA process,and results principally from the presence of osteoclasts at theinterface of bone and pannus. Osteoclasts are multinucleated cells that,upon adhesion to the bone tissue, form a closed compartment, withinwhich the osteoclasts secrete proteases (Cathepsin K, MMP9) that degradethe bone tissue. The osteoclast population in the joint is abnormallyincreased by osteoblast formation from precursor cells induced by thesecretion of the receptor activator of NFκB ligand (RANKL) by activatedSFs and T-cells.

Various collagen types have a key role in defining the stability of theextracellular matrix (ECM). Collagens type I and collagen type II, forexample, are the main components of bone and cartilage, respectively.Collagen proteins typically organise into multimeric structures referredto as collagen fibrils. Native collagen fibrils are very resistant toproteolytic cleavage. Only a few types of ECM-degrading proteins havebeen reported to have the capacity to degrade native collagen: MMPs andCathepsins. Among the Cathepsins, cathepsin K, which is active mainly inosteoclasts, is the best characterised. Among the MMPs, MMP1, MMP2, MMP8MMP13 and MMP14 are known to have collagenolytic properties. Thecorrelation between an increased expression of MMP1 by synovialfibroblasts (SFs) and the progression of the arthritic disease iswell-established and is predictive for joint erosive processes (Cunnaneet al., 2001). In the context of RA, therefore, MMP1 represents a highlyrelevant collagen degrading protein. In vitro, the treatment of culturedSFs with cytokines relevant in the RA pathology (e.g. TNF-α and IL1β)will increase the expression of MMP1 by these cells (Andreakos et al.,2003). Monitoring the levels of MMP1 expressed by SFs therefore is arelevant readout in the field of RA as it is indicative for theactivation of SFs towards an erosive phenotype that, in vivo, isresponsible for cartilage degradation. Inhibition of the MMP1 expressionby SFs represents a valuable therapeutic approach towards the treatmentof RA.

The activity of the ECM-degrading proteins can also be causative orcorrelate with the progression of various diseases different from RA, ase.g. other diseases that involve the degradation of the joints. Thesediseases include, but are not limited to, psoriatic arthritis, juvenilearthritis, early arthritis, reactive arthritis, osteoarthritis, andankylosing spondylitis. Other diseases that may be treatable withcompounds identified according to the present invention and using thetargets involved in the expression of MMPs as described herein areosteoporosis, muscular skeletal diseases like tendonitis and periodontaldisease (Gapski et al., 2004), cancer metastasis (Coussens et al.,2002), airway diseases (COPD, asthma) (Suzuki et al., 2004), lung, renalfibrosis (Schanstra et al., 2002), liver fibrosis associated withchronic hepatitis C (Reiff et al., 2005), cardio-vascular diseases likeatherosclerosis and heart failure (Creemers et al., 2001), andneurological diseases like neuroinflammation and multiple sclerosis(Rosenberg, 2002). Patients suffering from such diseases may benefitfrom stabilizing the ECM (by protecting it from degradation).

The 471-amino acid serine/threonine kinase identified asMitogen-Activated Protein Kinase-Activated Protein Kinase 5 (MAPKAPK5 orPRAK) is expressed in a wide panel of tissues. The protein contains itscatalytic domain at the N-terminal end and both a nuclear localizationsignal (NLS) and nuclear export signal (NES) at its C-terminal end.Endogenous MAPKAPK5 is predominantly present in the cytoplasm, butstress or cytokine activation of the cells mediates its translocationinto the nucleus (New et al., 2003). This event is dependent onphosphorylation of MAPKAPK5. Thr182 is the regulatory phosphorylationsite of MAPKAPK5. Although the p38α kinase is able to phosphorylateMAPKAPK5 in an overexpression setting, experiments with endogenousMAPKAPK5 do not support this hypothesis (Shi et al., 2003). MAPKAPK5knock-out mice have been generated that are viable and fertile. Thephenotype of these mice is quite different from that of mice deficientfor MAPKAPK2, a MAPKAPK5 related kinase that is regulated by p38α (Shiet al., 2003). This indicates that the function of each protein isdistinct and that neither kinase can compensate for the other'sactivity. Taken together, MAPKAPK5 and MAPKAPK2 represent distincttargets with a non-redundant role. MAPK6 (also referred to as ERK3) hasrecently been identified as a physiologically relevant substrate forMAPKAPK5, defining a novel signal transduction pathway (Seternes et al.,2004).

BACKGROUND OF THE INVENTION

NSAIDS (Non-steroidal anti-inflammatory drugs) are used to reduce thepain associated with RA and improve life quality of the patients. Thesedrugs will not, however, put a brake on the RA-associated jointdestruction.

Corticosteroids were found to decrease the progression of RA as detectedradiographically and are used at low doses to treat part of the RApatients (30 to 60%). Serious side effects, however, are associated withlong corticosteroid use (Skin thinning, osteoporosis, cataracts,hypertension, hyperlipidemia).

Synthetic DMARDs (Disease-Modifying Anti-Rheumatic Drugs) (e.g.methotrexate, leflunomide, sulfasalazine) mainly tackle theimmuno-inflammatory component of RA. As a main disadvantage, these drugsonly have a limited efficacy (joint destruction is only slowed down butnot blocked by DMARDs such that disease progression in the long termcontinues). The lack of efficacy is indicated by the fact that, onaverage, only 30% of the patients achieve an ACR40 score after 24 monthstreatment with methotrexate. This means that, according to the AmericanCollege of Rheumatology, only 30% of the patients do achieve a 50%improvement of their symptoms (O'Dell et al., 1996). In addition, theprecise mechanism of action of DMARDs is often unclear.

Biological DMARDs (Infliximab, Etanercept, Adalimumab, Rituximab,CTLA4-Ig) are therapeutic proteins that do inactivate cytokines (e.g.TNF-α) or cells (e.g. T-cells or B-cells) that have an important role inthe RA pathophysiology (Kremer et al., 2003; Edwards et al., 2004).Although the TNF-α-blockers (Infliximab, Etanercept, Adalimumab) andmethotrexate combination therapy is the most effective RA treatmentcurrently available, it is striking that even this therapy only achievesa 50% improvement (ACR40) in disease symptoms in 50-60% of patientsafter 12 months therapy (St Clair et al., 2004). Some adverse eventswarnings for anti-TNF-α drugs exist, shedding a light on the sideeffects associated to this type of drugs. Increased risk for infections(tuberculosis) hematologic events and demyelinating disorders have beendescribed for the TNF-α blockers (see also Gomez-Reino et al., 2003).Besides the serious side effects, the TNF-α blockers do also share thegeneral disadvantages of the biological class of therapeutics, which arethe unpleasant way of administration (frequent injections accompanied byinfusion site reactions) and the high production cost. Newer agents inlate development phase target T-cell co-stimulatory molecules andB-cells. The efficacy of these agents is expected to be similar to thatof the TNF-α blockers. The fact that a variety of targeted therapieshave similar but limited efficacies, suggests that there is amultiplicity of pathogenic factors for RA. This is also indicative forthe deficiencies in our understanding of pathogenic events relevant toRA.

The current therapies for RA are not satisfactory due to a limitedefficacy (No adequate therapy exists for 30% of the patients). Thiscalls for additional strategies to achieve remission. Remission isrequired since residual disease bears the risk of progressive jointdamage and thus progressive disability. Inhibiting theimmuno-inflammatory component of the RA disease, which represents themain target of drugs currently used for RA treatment, does not result ina blockade of joint degradation, the major hallmark of the disease.

US 2005/0009832 describes substitutedimidazolo[1,2-a]pyrazine-8-yl-amines as modulators of protein kinases,including MAPKAPK5. W002/056888 describes inhibitors of MAPKAPK5 as TNFmodulators able to regulate the expression of certain cytokines. Neitherof these prior art references discloses any compound within the scope ofthe class of compounds described herein below.

SUMMARY OF THE INVENTION

The present invention is based on the discovery of that MAPKAPK5functions in the pathway that results in the expression of MMP1, andthat inhibitors of MAPKAPK5 activity, such as the compounds of thepresent invention, are useful for the treatment of diseases involvingthe abnormally high expression of MMP activity.

The present matrix metallo proteinase inhibiting compounds of thepresent invention may be described generally asimidazo[1,2-a]pyrazine-8-yl-amines substituted in the 5-position by anaromatic group capable of donating electrons to, and an 8-aminosubstituent capable of accepting electrons from, theimidazo[12.a]pyrazine ring. In particular, the 5-substituent group ischaracterized as having a hydrogen bond donor-acceptor functionality,whereas the substituent on the 8-amino group must be sufficientlyelectron-withdrawing to polarise the N-H bond of the 8-substituent, oralternatively, the 8-NH group is capable of participating inpi-conjugation with the substituent group on the 8-NH group.

The compounds of the present invention may show less toxicity, goodabsorption, good half-life, good solubility, low protein bindingaffinity, less drug-drug interaction, and good metabolic stability.

More particularly, the present invention relates to compounds havingmatrix metallo proteinase inhibiting properties in a mammalian cell,according to formula (I):

wherein:

-   A and B are independently CR4R″, NR″, oxygen or sulfur;-   AA is CR4 or N;-   D is C═O, CR4R″ or NR″;-   E is NH or CR″R6, when k is zero, and is NH or CR″R6a, when k is    one;-   F is sulfur, oxygen or NH;-   T is oxygen or NR;-   U, V, W and X are independently CR″R7 or NR″;-   Y is CR″ or N;-   Z is hydrogen, amino, hydroxyl, lower alkoxy, carbamoyl, carboxyl,    SO₂Rz, SO₂NRRz, —NR(CO)(CH2)d-Rz, —NRRz, —(CO)—ORz,    —(CO)—NR(CH2)d-Rz, or

-   R is independently hydrogen or lower alkyl;-   R′ is independently hydrogen or lower alkyl;-   R″ is H or forms a double bond with an adjacent atom;-   R1 is H; R4; or lower alkyl, lower cycloalkyl and lower alkyl-lower    cycloalkyl, optionally substituted with one or more R4;-   R2 is H; lower alkyl, lower cycloalkyl and lower alkyl-lower    cycloalkyl, optionally substituted with one or more of F and Cl;-   R3 is H or forms a double bond with an adjacent R″;-   R4 is H, F, Cl; CN; COOR5; OR5; C(O)N(R5R5a); S(O)₂N(R5R5a); lower    alkyl; O— lower NH-lower alkyl; S-lower alkyl; COO— lower alkyl;    OC(O)— lower alkyl; C(O)N(R5)-lower alkyl; S(O)₂N(R5)-lower alkyl;    S(O)N(R5)-lower alkyl; S(O)₂-lower alkyl; S(O)-lower alkyl;    N(R5)S(O)₂-lower alkyl; and N(R5)S(O)-lower alkyl; wherein each    lower alkyl is optionally substituted with one or more of F and Cl;-   R5 and R5a are independently    -   H; F, Cl; or lower alkyl, lower cycloalkyl, or lower alkyl-lower        cycloalkyl optionally substituted with one or more of F and Cl;-   R6 is hydrogen, amino, hydroxyl, carbamoyl, carboxyl, SO₂R, NRR′,    —(CO)—OR, or —CO)—NRR′;-   R6a is R6, Cl, F, lower alkoxy, cyano, trifluoromethoxy; or together    with the adjacent be —(CHR″)_(n)—NR—(CHR″)_(p)—, and form a five or    six member heterocyclic ring fused to the ring to which they are    bonded;-   R7 is independently hydrogen, halogen, lower alkyl or lower alkoxy;-   Rz is hydrogen, lower alkyl, lower alkanoyl, phenyl, 1-lower alkyl    pyrrolidin-3-yl, pyrazol-4-yl, pyrazol-2-yl, or lower alkyl, lower    alkanoyl, phenyl, 1-loweralkyl pyrrolidin-3-yl, pyrazol-4-yl,    pyrazol-2-yl or pyrid-3-yl substituted by one or more of hydroxyl,    amino, mono- or di-loweralkylamino, acetamidyl, lower alkanoyl,    lower alkyl, 4-hydroxy-phenyl, 3-aminomethylphenyl, lower alkyl    sulfonyl, 4-diloweralkylaminophenyl, pyrid-3-yl, 1H-indol-3-yl,    morpholin-4-yl,;-   R and Rz together may be —(CHR)_(q)-T-(CHR)_(r)— and form a five or    six member heterocyclic ring with the nitrogen to which they are    bonded;-   Rz and R7 together may be —(CHR″)_(n)—NR—(CHR″)_(p)—, and form a    five or six member heterocyclic ring fused to the ring to which they    are bonded;-   b and d are independently 0 or 1; provided at least one of b or d is    1;-   k is 0 or 1;-   m is 0 or 1;-   n and p are independently 0, 1 or 2;-   q and r are 1 or 2;-   x is o or 1;-   with the provisos that: (1) when m is zero, and (a) either U or W is    NR″, then Z is not carbamoyl; and    -   (b) x is land U is NR″, then W is not NR″;    -   (2) (a) when m is 1, or (b) when U, V, W, X and Y form phenyl        and X is C-lower alkoxy, or (c) when Rz together with R7 form        indolyl, then R6 is not carbamoyl; and    -   (3) at least one of R7 is other than hydrogen;        or a pharmaceutically acceptable salt, hydrate, solvate or        prodrug thereof.

A preferred aspect of the present invention is a subclass of compoundsaccording to formula II,

wherein

-   A and B are independently CR4R″, NR″, oxygen or sulfur;-   AA is CR4 or N;-   D is C═O, CR4R″ or NR″;-   E is NH or CR″R6, when k is zero, and is NH or CR″R6a, when k is    one;-   F is sulfur, oxygen or NH;-   T is oxygen or NR;-   R″ is H or forms a double bond with an adjacent atom;-   R is independently hydrogen or lower alkyl;-   R′ is independently hydrogen or lower alkyl;-   R1 is H; R4; or lower alkyl, lower cycloalkyl and lower alkyl-lower    cycloalkyl, optionally substituted with one or more R4;-   R2 is H; lower alkyl, lower cycloalkyl and lower alkyl-lower    cycloalkyl, optionally substituted with one or more of F and Cl;-   R4 is H, F, Cl; CN; COOR5; OR5; C(O)N(R5R5a); S(O)₂N(R5R5a); lower    alkyl; O— -lower alkyl NH-lower alkyl; S-lower alkyl; COO— lower    alkyl; OC(O)— lower alkyl; C(O)N(R5)— lower alkyl; S(O)₂N(R5)-lower    alkyl; S(O)N(R5)-lower alkyl; S(O)₂-lower alkyl; S(O)-lower alkyl;    N(R5)S(O)₂-lower alkyl; and N(R5)S(O)-lower alkyl; wherein each    lower alkyl is optionally substituted with one or more of F and Cl;-   R5 and R5a are independently    -   H; F, Cl; or lower alkyl, lower cycloalkyl, or lower alkyl-lower        cycloalkyl optionally substituted with one or more of F and Cl;-   R6 is hydrogen, amino, hydroxyl, carbamoyl, carboxyl, SO₂R, NRR′,    —(CO)—OR, or —(CO)—NRR′;-   R6a is R6, Cl, F, lower alkoxy, cyano, trifluoromethoxy; or together    with the adjacent be —(CHR″)_(n)—NR—(CHR″)_(p)—, and form a five or    six member heterocyclic ring fused to the ring to which they are    bonded;-   R8 is phenyl independently substituted by R_(a) in the    ortho-position, by R_(b) in the meta-position, and by R_(c) in the    para-position; pyrid-3-yl; pyrid-3-yl substituted by R_(c) in the    5-position; or cyclohexyl independently substituted by R_(a) in the    2-position, and by R_(d) in the 4-position;    -   R_(a) is hydrogen, halogen, lower alkyl, trifluoromethyl or        lower alkoxy;    -   R_(b) is hydrogen, trifluoromethyl, lower alkoxy, lower alkyl        sulfonamide, carboxyl, —NR_(e)R_(f), —(CO)—OR, or        —(CO)—NR_(e)R_(f);    -   R_(c) is hydrogen, amino, hydroxyl, lower alkoxy, carbamoyl,        carboxyl, SO₂R, SO₂NR_(e)R_(f), NR_(e)R_(f), —(CO)—OR, or        —(CO)—NR_(e)R_(f); or    -   R_(b) and R_(c) can together form a benzdiazole, or indole        substituted in the 3-position by R′;    -   R_(d) is hydroxyl, halogen, amino, lower alkoxy, or NR_(e)R_(f);    -   R_(e) and R_(f) are independently hydrogen, 1-loweralkyl        pyrrolidin-3-yl, 1-R-pyrazol-4-yl, lower alkanoyl, phenyl, or        lower alkyl optionally substituted by one or more of        4-hydroxy-phenyl, 3-aminomethylphenyl, lower alkyl sulfonyl,        4-diloweralkylaminophenyl, pyrid-3-yl, 1H-indol-3-yl,        morpholin-4-yl, hydroxyl, amino, mono- or di-loweralkylamino, or        by lower alkanoyl; or R′ and R″ together are        —(CHR)_(n)-T-(CHR)_(n)— and form a five or six member        heterocyclic ring with the nitrogen to which they are bonded;-   m is 0, 1, or2;-   n is 1 or 2;-   with the proviso that (1) (a) when R8 is pyridy-3-yl, or (b) when    R_(b) is lower alkoxy or (c) together with R_(c) are indolyl, then    R_(e) is not carbamoyl;    -   (2) when R_(e) is hydroxyl, then R_(g) is not lower alkyl; and    -   (3) at least one of R_(a), R_(b) and R_(c) is other than        hydrogen;        or a pharmaceutically acceptable salt, hydrate, solvate or        prodrug thereof.

Another aspect of the present invention is compounds according toformula III:

wherein

R¹ is H, or substituted or unsubstituted alkyl; R² is H, lower alkyl,lower cycloalkyl and lower alkyl-lower cycloalkyl, optionallysubstituted with one or more of F and Cl; R⁸ is selected fromsubstituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl, substituted orunsubstituted pyridyl, substituted or unsubstituted pyrimidine, andsubstituted or unsubstituted pyrazine, substituted or unsubstitutedpyrrole, substituted or unsubstituted pyrazole and substituted orunsubstituted imidazole; and R⁹ is selected from substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, substituted or unsubstitutedheteroaryl; or a pharmaceutically acceptable salt, solvate or prodrugthereof; and stereoisomers, isotopic variants and tautomers thereof.

In one embodiment, with respect to compounds of formula III, R¹ is H,Me, Et, i-Pr or CF₃.

In one embodiment, with respect to compounds of formula III, R¹ is H.

In one embodiment, with respect to compounds of formula III, R² is H,Me, Et, i-Pr or CF₃.

In one embodiment, with respect to compounds of formula III, R² is H.

Another aspect of the present invention relates to compounds accordingto formula IVa, IVb, IVc, or IVd:

and wherein L is a bond, —CO—, SO₂, —(CH₂)_(m1)—, —O(CH₂)_(m1)—,—NH(CH₂)_(m1)—, —CON(H)(CH₂)_(m1)—, or —SO₂NH(CH₂)_(m1)—; the subscriptm1 is selected from 1-4; the ring P is substituted or unsubstitutedheterocycloalkyl; the subscript n is selected from 1-4; each R^(8a) isindependently selected from hydrogen, substituted or unsubstitutedalkyl, alkoxy, cyano, carbamoyl, CHO, and halo; and R⁹ is independentlyselected from substituted or unsubstituted aryl and heteroaryl; or apharmaceutically acceptable salt, solvate or prodrug thereof; andstereoisomers, isotopic variants and tautomers thereof.

In a one embodiment, with respect to compounds of formulae III-IVd, R⁹is substituted or unsubstituted aryl. In another embodiment, R⁹ issubstituted or unsubstituted phenyl.

In a one embodiment, with respect to compounds of formulae III-IVd, R⁹is substituted or unsubstituted heteroaryl. In another embodiment, R⁹ issubstituted or unsubstituted pyridyl.

In a one embodiment, with respect to compounds of formulae III-IVd, R⁹is selected from substituted or unsubstituted phenyl, indolyl,isoinolyl, pyrrolyl, furanyl, thienyl, pyrazolyl, oxazolyl, andthiazolyl.

In a further aspect, the present invention provides pharmaceuticalcompositions comprising an imidazolopyrazine compound of the invention,and a pharmaceutical carrier, excipient or diluent. In this aspect ofthe invention, the pharmaceutical composition can comprise one or moreof the compounds described herein. Moreover, the compounds of thepresent invention useful in the pharmaceutical compositions andtreatment methods disclosed herein, are all pharmaceutically acceptableas prepared and used.

Another aspect of this invention relates to the use of the presentcompound in a therapeutic method, a pharmaceutical composition, and themanufacture of such composition, useful for the treatment of diseasesinvolving inflammation, collagen degradation, and in particular,diseases characteristic of abnormal matrix metallo protease (MMP1)and/or Mitogen-Activated Protein-Kinase Activated Protein Kinase 5(MAPKAPK5) activity, of which rheumatoid arthritis (RA) is a particularsuch disease. This invention also relates to processes for thepreparation of the present compounds.

Other objects and advantages will become apparent to those skilled inthe art from a consideration of the ensuing detailed description,considered in conjunction with the following illustrative drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. This diagram shows the striking histological differences betweena healthy joint and that of a RA patient ((From Smolen and Steiner,2003).

FIG. 2. This chart shows the increased expression of MMP1 in synovialfibroblasts triggered with cytokines involved in rheumatoid arthritispathology.

FIG. 3. This graph shows the dose-dependent inhibition of the“TNF-α-based trigger”-induced expression of MMP1 by SFs by a knownanti-inflammatory compound.

FIG. 4. This gel shows the reduction, at the protein level, of theexpression of MAPKAPK5 in SFs by infection of the cells with Ad-siRNAvirus targeting MAPKAPK5.

FIG. 5. This chart shows the reduction of ‘complex trigger’ inducedlevels of MMP1 expression by SFs by an Ad-siRNA virus targetingMAPKAPK5.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

When describing the compounds, pharmaceutical compositions containingsuch compounds and methods of using such compounds and compositions, thefollowing terms have the following meanings unless otherwise indicated.It should also be understood that any of the moieties defined forthbelow may be substituted with a variety of substituents, and that therespective definitions are intended to include such substituted moietieswithin their scope. By way of non-limiting example, such substituentsmay include e.g. halo (such as fluoro, chloro, bromo), —CN, —CF₃, —OH,—OCF₃, C₂-C₆ alkenyl, C₃-C₆ alkynyl, C₁-C₆ alkoxy, aryl and di-C₁-C₆alkylamino. It should be further understood that the terms “groups” and“radicals” can be considered interchangeable when used herein.

“Alkoxy” means alkyl-O—. Exemplary alkoxy includes methoxy, ethoxy,n-propoxy, i-propoxy, n-butoxy, and heptoxy. Preferred alkoxy groups arelower alkoxy.

“Alkyl” means straight or branched aliphatic hydrocarbon having 1 toabout 20 carbon atoms. Preferred alkyl has 1 to about 12 carbon atoms.More preferred is lower alkyl. Branched means that one or more loweralkyl groups such as methyl, ethyl or propyl is attached to a linearalkyl chain.

“Alkyl amino” means alkyl-NH—. Preferred alkyl amino is (C₁-C₆)-alkylamino. Exemplary alkyl amino includes methylamino and ethylamino.

“Amino lower alkanoyl” means NH₂—R—CO—, where R is lower alkylene.Preferred groups include aminoethanoyl and aminoacetyl.

“Carbamoyl lower alkyl” means the radical NH₂CO-lower alkyl-. Preferredgroups include carbamoylethyl and carbamoylmethyl.

“Carboxy lower alkyl ester” means a lower alkyl ester of a carboxyradical, —COO— group.

“Compounds of the present invention”, and equivalent expressions, aremeant to embrace compounds of Formula (I, II or III) as hereinbeforedescribed, which expression includes the prodrugs, the pharmaceuticallyacceptable salts, and the solvates, e.g., hydrates, where the context sopermits. Similarly, reference to intermediates, whether or not theythemselves are claimed, is meant to embrace their salts, and solvates,where the context so permits.

“Expression” means endogenous expression.

“Halo” or “halogen” means fluoro, chloro, bromo, or iodo.

“Hydrogen” means in the context of a substituent that —H is present atthe compound position and also includes its isotope, deuterium.

“Lower alkanoyl amino” means an amino group with an organic functionalgroup R—CO—, where R represents a lower alkyl group.

“Lower alkyl” means 1 to about 6 carbon atoms in a linear alkyl chainthat may be straight or branched.

“Lower alkoxy” means 1 to about 6 carbon atoms in a linear alkyl chainthat may be straight or branched, and that is bonded by an oxygen atom.

“Lower alkyl sulphonamide” refers to a lower alkyl amide of sulphonamideof the formula —SO₂NR*R*, where R* is hydrogen or lower alkyl, and atleast one R* is lower alkyl.

“Prophylaxis” means a measure taken for the prevention of a disease.

“Solvate” means a physical association of a compound useful in thisinvention with one or more solvent molecules. This physical associationincludes hydrogen bonding. In certain instances the solvate will becapable of isolation, for example when one or more solvent molecules areincorporated in the crystal lattice of the crystalline solid. “Solvate”encompasses both solution-phase and isolable solvates. The compounds ofthe invention may be prepared e.g. in crystalline form and may besolvated or hydrated. Suitable solvates include pharmaceuticallyacceptable solvates, such as hydrates, and further include bothstoichiometric solvates and non-stoichiometric solvates. Conventionalsolvents include water, ethanol, acetic acid and the like, therefore,representative solvates include hydrates, ethanolates and methanolates.

“Substituted” means that one atom or group of atoms in a molecule isreplaced by another atom or group.

“Sulphonamide” refers to a group of compounds containing the chemicalgroup —SO₂NH₂.

“Therapeutically effective amount” means that amount of a drug orpharmaceutical agent that will elicit the biological or medical responseof a subject that is being sought by a medical doctor or otherclinician. The “therapeutically effective amount” can vary depending onthe compound, the disease and its severity, and the age, weight, etc.,of the subject to be treated. In particular, with regard to treating andisease condition characterized by the degradation of extracellularmatrix, the term “effective matrix metallo-protease inhibiting amount”is intended to mean that effective amount of an compound of the presentinvention that will bring about a biologically meaningful decrease inthe production of MMP-1 in the subject's disease affected tissues suchthat extracellular matrix degradation is meaningfully reduced. Acompound having matrix metallo-protease inhibiting properties or a“matrix metallo-protease inhibiting compound” means a compound of thepresent invention that provided to a cell in effective amounts is ableto cause a biologically meaningful decrease in the production of MMP-1in such cells.

“Aryl” refers to a monovalent aromatic hydrocarbon group derived by theremoval of one hydrogen atom from a single carbon atom of a parentaromatic ring system. Typical aryl groups include, but are not limitedto, groups derived from aceanthrylene, acenaphthylene,acephenanthrylene, anthracene, azulene, benzene, chrysene, coronene,fluoranthene, fluorene, hexacene, hexaphene, hexalene, as-indacene,s-indacene, indane, indene, naphthalene, octacene, octaphene, octalene,ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene,phenalene, phenanthrene, picene, pleiadene, pyrene, pyranthrene,rubicene, triphenylene, trinaphthalene and the like. Particularly, anaryl group comprises from 6 to 14 carbon atoms.

“Substituted Aryl” includes those groups recited in the definition of“substituted” herein, and particularly refers to an aryl group that mayoptionally be substituted with 1 or more substituents, for instance from1 to 5 substituents, particularly 1 to 3 substituents, selected from thegroup consisting of acyl, acylamino, acyloxy, alkenyl, substitutedalkenyl, alkoxy, substituted alkoxy, alkoxycarbonyl, alkyl, substitutedalkyl, alkynyl, substituted alkynyl, amino, substituted amino,aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy,azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen,hydroxyl, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thiol,alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—.

“Bicycloaryl” refers to a monovalent aromatic hydrocarbon group derivedby the removal of one hydrogen atom from a single carbon atom of aparent bicycloaromatic ring system. Typical bicycloaryl groups include,but are not limited to, groups derived from indane, indene, naphthalene,tetrahydronaphthalene, and the like. Particularly, an aryl groupcomprises from 8 to 11 carbon atoms.

“Bicycloheteroaryl” refers to a monovalent bicycloheteroaromatic groupderived by the removal of one hydrogen atom from a single atom of aparent bicycloheteroaromatic ring system. Typical bicycloheteroarylgroups include, but are not limited to, groups derived from benzofuran,benzimidazole, benzindazole, benzdioxane, chromene, chromane, cinnoline,phthalazine, indole, indoline, indolizine, isobenzofuran, isochromene,isoindole, isoindoline, isoquinoline, benzothiazole, benzoxazole,naphthyridine, benzoxadiazole, pteridine, purine, benzopyran,benzpyrazine, pyridopyrimidine, quinazoline, quinoline, quinolizine,quinoxaline, benzomorphan, tetrahydroisoquinoline, tetrahydroquinoline,and the like. Preferably, the bicycloheteroaryl group is between 9-11membered bicycloheteroaryl, with 5-10 membered heteroaryl beingparticularly preferred. Particular bicycloheteroaryl groups are thosederived from benzothiophene, benzofuran, benzothiazole, indole,quinoline, isoquinoline, benzimidazole, benzoxazole and benzdioxane.

“Carbamoyl” refers to the radical —C(O)N(R⁴²)₂ where each R⁴² group isindependently hydrogen, alkyl, cycloalkyl or aryl, as defined herein,which may be optionally substituted as defined herein. In a specificembodiment, the term “carbamoyl” refers to —C(O)—NH₂.

“Cycloalkyl” refers to cyclic hydrocarbyl groups having from 3 to about10 carbon atoms and having a single cyclic ring or multiple condensedrings, including fused and bridged ring systems, which optionally can besubstituted with from 1 to 3 alkyl groups. Such cycloalkyl groupsinclude, by way of example, single ring structures such as cyclopropyl,cyclobutyl, cyclopentyl, cyclooctyl, 1-methylcyclopropyl,2-methylcyclopentyl, 2-methylcyclooctyl, and the like, and multiple ringstructures such as adamantanyl, and the like.

“Substituted cycloalkyl” includes those groups recited in the definitionof “substituted” herein, and particularly refers to a cycloalkyl grouphaving 1 or more substituents, for instance from 1 to 5 substituents,and particularly from 1 to 3 substituents, selected from the groupconsisting of acyl, acylamino, acyloxy, alkoxy, substituted alkoxy,alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino,aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy,azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen,hydroxyl, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy,thioketo, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—.

“Substituted” refers to a group in which one or more hydrogen atoms areeach independently replaced with the same or different substituent(s).Typical substituents include, but are not limited to, —X, —R⁴⁶, —O⁻,═O,—OR⁴⁶, —SR⁴⁶, —S⁻, ═S, —NR⁴⁶R⁴⁷, ═NR⁴⁶, —CX₃, —CF₃, —CN, —OCN, —SCN,—NO, —NO₂, ═N₂, —N₃, —S(O)₂O⁻, —S(O)₂OH, —S(O)₂R⁴⁶, —OS(O₂)O⁻,—OS(O)₂R⁴⁶,—P(O)(O⁻)₂, —P(O)(OR⁴⁶)(O⁻), —OP(O)(OR⁴⁶)(OR⁴⁷), —C(O)R⁴⁶,—C(S)R⁴⁶, —C(O)OR⁴⁶, —C(O)NR⁴⁶R⁴⁷, —C(O)O⁻, —C(S)OR⁴⁶, —NR⁴⁸C(O)NR⁴⁶R⁴⁷,—NR⁴⁸C(S)NR⁴⁶R⁴⁷, —NR⁴⁹C(NR⁴⁸)NR⁴⁶R⁴⁷ and —C(NR⁴⁸)NR⁴⁶R⁴⁷, where each Xis independently a halogen; each R⁴⁶, R⁴⁷, R⁴⁸ and R⁴⁹ are independentlyhydrogen, alkyl, substituted alkyl, aryl, substituted alkyl, arylalkyl,substituted alkyl, cycloalkyl, substituted alkyl, cycloheteroalkyl,substituted cycloheteroalkyl, heteroalkyl, substituted heteroalkyl,heteroaryl, substituted heteroaryl, heteroarylalkyl, substitutedheteroarylalkyl, —NR⁵⁰R⁵¹, —C(O)R⁵⁰ or —S(O)₂R⁵⁰ or optionally R⁵⁰ andR⁵¹ together with the atom to which they are both attached form acycloheteroalkyl or substituted cycloheteroalkyl ring; and R⁵⁰ and R⁵¹are independently hydrogen, alkyl, substituted alkyl, aryl, substitutedalkyl, arylalkyl, substituted alkyl, cycloalkyl, substituted alkyl,cycloheteroalkyl, substituted cycloheteroalkyl, heteroalkyl, substitutedheteroalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl orsubstituted heteroarylalkyl.

Examples of representative substituted aryls include the following

In these formulae one of R⁵² and R⁵³ may be hydrogen and at least one ofR⁵² and R⁵³ is each independently selected from alkyl, alkenyl, alkynyl,cycloheteroalkyl, alkanoyl, alkoxy, aryloxy, heteroaryloxy, alkylamino,arylamino, heteroarylamino, NR⁵⁴COR⁵⁵, NR⁵⁴SOR⁵⁵, NR⁵⁴SO₂R⁵⁷, COOalkyl,COOaryl, CONR⁵⁴R⁵⁵, CONR⁵⁴OR⁵⁵, NR⁵⁴R⁵⁵, SO₂NR⁵⁴R⁵⁵, S-alkyl, S-alkyl,SOalkyl, SO₂alkyl, Saryl, SOaryl, SO₂aryl; or R⁵² and R⁵³ may be joinedto form a cyclic ring (saturated or unsaturated) from 5 to 8 atoms,optionally containing one or more heteroatoms selected from the group N,O or S. R⁵⁴, R⁵⁵, and R⁵⁶ are independently hydrogen, alkyl, alkenyl,alkynyl, perfluoroalkyl, cycloalkyl, cycloheteroalkyl, aryl, substitutedaryl, heteroaryl, substituted or hetero alkyl or the like.

“Hetero” when used to describe a compound or a group present on acompound means that one or more carbon atoms in the compound or grouphave been replaced by a nitrogen, oxygen, or sulfur heteroatom. Heteromay be applied to any of the hydrocarbyl groups described above such asalkyl, e.g. heteroalkyl, cycloalkyl, e.g. cycloheteroalkyl, aryl, e.g.heteroaryl, cycloalkenyl, cycloheteroalkenyl, and the like having from 1to 5, and especially from 1 to 3 heteroatoms.

“Heteroaryl” refers to a monovalent heteroaromatic group derived by theremoval of one hydrogen atom from a single atom of a parentheteroaromatic ring system. Typical heteroaryl groups include, but arenot limited to, groups derived from acridine, arsindole, carbazole,β-carboline, chromane, chromene, cinnoline, furan, imidazole, indazole,indole, indoline, indolizine, isobenzofuran, isochromene, isoindole,isoindoline, isoquinoline, isothiazole, isoxazole, naphthyridine,oxadiazole, oxazole, perimidine, phenanthridine, phenanthroline,phenazine, phthalazine, pteridine, purine, pyran, pyrazine, pyrazole,pyridazine, pyridine, pyrimidine, pyrrole, pyrrolizine, quinazoline,quinoline, quinolizine, quinoxaline, tetrazole, thiadiazole, thiazole,thiophene, triazole, xanthene, and the like. Preferably, the heteroarylgroup is between 5-15 membered heteroaryl, with 5-10 membered heteroarylbeing particularly preferred. Particular heteroaryl groups are thosederived from thiophene, pyrrole, benzothiophene, benzofuran, indole,pyridine, quinoline, imidazole, oxazole and pyrazine.

Examples of representative heteroaryls include the following:

wherein each Y is selected from carbonyl, N, NR⁵⁸, O, and S; and R⁵⁸ isindependently hydrogen, alkyl, cycloalkyl, cycloheteroalkyl, aryl,heteroaryl, heteroalkyl or the like.

As used herein, the term “cycloheteroalkyl” refers to a stableheterocyclic non-aromatic ring and fused rings containing one or moreheteroatoms independently selected from N, O and S. A fused heterocyclicring system may include carbocyclic rings and need only include oneheterocyclic ring. Examples of heterocyclic rings include, but are notlimited to, piperazinyl, homopiperazinyl, piperidinyl and morpholinyl,and are shown in the following illustrative examples:

wherein each X is selected from CR⁵⁸ ₂, NR⁵⁸, O and S; and each Y isselected from NR⁵⁸, O and S; and R⁵⁸ is independently hydrogen, alkyl,cycloalkyl, cycloheteroalkyl, aryl, heteroaryl, heteroalkyl or the like.These cycloheteroalkyl rings may be optionally substituted with one ormore groups selected from the group consisting of acyl, acylamino,acyloxy, alkoxy, substituted alkoxy, alkoxycarbonyl,alkoxycarbonylamino, amino, substituted amino, aminocarbonyl,aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy, azido, carboxyl,cyano, cycloalkyl, substituted cycloalkyl, halogen, hydroxyl, keto,nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioketo, thiol,alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—. Substitutinggroups include carbonyl or thiocarbonyl which provide, for example,lactam and urea derivatives.

Examples of representative cycloheteroalkenyls include the following:

wherein each X is selected from CR⁵⁸ ₂, NR⁵⁸, O and S; and each Y isselected from carbonyl, N, NR⁵⁸, O and S; and R⁵⁸ is independentlyhydrogen, alkyl, cycloalkyl, cycloheteroalkyl, aryl, heteroaryl,heteroalkyl or the like.

Examples of representative aryl having hetero atoms containingsubstitution include the following:

wherein each X is selected from C—R⁵⁸ ₂, NR⁵⁸, O and S; and each Y isselected from carbonyl, NR⁵⁸, O and S; and R⁵⁸ is independentlyhydrogen, alkyl, cycloalkyl, cycloheteroalkyl, aryl, heteroaryl,heteroalkyl or the like.

One having ordinary skill in the art of organic synthesis will recognizethat the maximum number of heteroatoms in a stable, chemically feasibleheterocyclic ring, whether it is aromatic or non aromatic, is determinedby the size of the ring, the degree of unsaturation and the valence ofthe heteroatoms. In general, a heterocyclic ring may have one to fourheteroatoms so long as the heteroaromatic ring is chemically feasibleand stable.

“Pharmaceutically acceptable” means approved by a regulatory agency ofthe Federal or a state government or listed in the U.S. Pharmacopoeia orother generally recognized pharmacopoeia for use in animals, and moreparticularly in humans.

“Pharmaceutically acceptable salt” refers to the non-toxic, inorganicand organic acid addition salts, and base addition salts, of compoundsof the present invention, in particular they are a salt of a compound ofthe invention that is pharmaceutically acceptable and that possesses thedesired pharmacological activity of the parent compound. These salts canbe prepared in situ during the final isolation and purification ofcompounds useful in the present invention. Such salts include: (1) acidaddition salts, formed with inorganic acids such as hydrochloric acid,hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and thelike; or formed with organic acids such as acetic acid, propionic acid,hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid,lactic acid, malonic acid, succinic acid, malic acid, maleic acid,fumaric acid, tartaric acid, citric acid, benzoic acid,3-(4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid,2-hydroxyethanesulfonic acid, benzenesulfonic acid,4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,4-toluenesulfonic acid, camphorsulfonic acid,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid,3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid,lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoicacid, salicylic acid, stearic acid, muconic acid, and the like; or (2)salts formed when an acidic proton present in the parent compound eitheris replaced by a metal ion, e.g., an alkali metal ion, an alkaline earthion, or an aluminum ion; or coordinates with an organic base such asethanolamine, diethanolamine, triethanolamine, N-methylglucamine and thelike. Salts further include, by way of example only, sodium, potassium,calcium, magnesium, ammonium, tetraalkylammonium, and the like; and whenthe compound contains a basic functionality, salts of non toxic organicor inorganic acids, such as hydrochloride, hydrobromide, tartrate,mesylate, acetate, maleate, oxalate and the like. The term“pharmaceutically acceptable cation” refers to a non toxic, acceptablecationic counter-ion of an acidic functional group. Such cations areexemplified by sodium, potassium, calcium, magnesium, ammonium,tetraalkylammonium cations, and the like.

“Pharmaceutically acceptable vehicle” refers to a diluent, adjuvant,excipient or carrier with which a compound of the invention isadministered.

“Preventing” or “prevention” refers to a reduction in risk of acquiringa disease or disorder (i.e., causing at least one of the clinicalsymptoms of the disease not to develop in a subject that may be exposedto or predisposed to the disease but does not yet experience or displaysymptoms of the disease).

“Prodrugs” refers to compounds, including derivatives of the compoundsof the invention, which have cleavable groups and become by solvolysisor under physiological conditions the compounds of the invention whichare pharmaceutically active in vivo. Such examples include, but are notlimited to, choline ester derivatives and the like, N-alkylmorpholineesters and the like.

“Subject” includes humans. The terms “human,” “patient” and “subject”are used interchangeably herein.

“Treating” or “treatment” of any disease or disorder refers, in oneembodiment, to ameliorating the disease or disorder (i.e., arresting orreducing the development of the disease or at least one of the clinicalsymptoms thereof). In another embodiment “treating” or “treatment”refers to ameliorating at least one physical parameter, which may not bediscernible by the subject. In yet another embodiment, “treating” or“treatment” refers to modulating the disease or disorder, eitherphysically, (e.g., stabilization of a discernible symptom),physiologically, (e.g., stabilization of a physical parameter), or both.In yet another embodiment, “treating” or “treatment” refers to delayingthe onset of the disease or disorder.

Other derivatives of the compounds of this invention have activity inboth their acid and acid derivative forms, but in the acid sensitiveform often offers advantages of solubility, tissue compatibility, ordelayed release in the mammalian organism (see, Bundgard, H., Design ofProdrugs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985). Prodrugs includeacid derivatives well know to practitioners of the art, such as, forexample, esters prepared by reaction of the parent acid with a suitablealcohol, or amides prepared by reaction of the parent acid compound witha substituted or unsubstituted amine, or acid anhydrides, or mixedanhydrides. Simple aliphatic or aromatic esters, amides and anhydridesderived from acidic groups pendant on the compounds of this inventionare preferred prodrugs. In some cases it is desirable to prepare doubleester type prodrugs such as (acyloxy)alkyl esters or((alkoxycarbonyl)oxy)alkylesters. Preferred are the C₁ to C₈ alkyl,C₂-C₈ alkenyl, aryl, C₇-C₁₂ substituted aryl, and C₇-C₁₂ arylalkylesters of the compounds of the invention.

As used herein, the term “isotopic variant” refers to a compound thatcontains unnatural proportions of isotopes at one or more of the atomsthat constitute such compound. For example, an “isotopic variant” of acompound can contain one or more non-radioactive isotopes, such as forexample, deuterium (²H or D), carbon-13 (¹³C), nitrogen-15 (¹⁵N), or thelike. It will be understood that, in a compound where such isotopicsubstitution is made, the following atoms, where present, may vary, sothat for example, any hydrogen may be ²H/D, any carbon may be ¹³C, orany nitrogen may be ¹⁵N, and that the presence and placement of suchatoms may be determined within the skill of the art. Likewise, theinvention may include the preparation of isotopic variants withradioisotopes, in the instance for example, where the resultingcompounds may be used for drug and/or substrate tissue distributionstudies. The radioactive isotopes tritium, i.e. ³H, and carbon-14, i.e.¹⁴C, are particularly useful for this purpose in view of their ease ofincorporation and ready means of detection. Further, compounds may beprepared that are substituted with positron emitting isotopes, such as¹¹C, ¹⁸F, ¹⁵O and ¹³N, and would be useful in Positron EmissionTopography (PET) studies for examining substrate receptor occupancy.

All isotopic variants of the compounds provided herein, radioactive ornot, are intended to be encompassed within the scope of the invention.

It is also to be understood that compounds that have the same molecularformula but differ in the nature or sequence of bonding of their atomsor the arrangement of their atoms in space are termed “isomers”. Isomersthat differ in the arrangement of their atoms in space are termed“stereoisomers”.

Stereoisomers that are not mirror images of one another are termed“diastereomers” and those that are non-superimposable mirror images ofeach other are termed “enantiomers”. When a compound has an asymmetriccenter, for example, it is bonded to four different groups, a pair ofenantiomers is possible. An enantiomer can be characterized by theabsolute configuration of its asymmetric center and is described by theR- and S-sequencing rules of Cahn and Prelog, or by the manner in whichthe molecule rotates the plane of polarized light and designated asdextrorotatory or levorotatory (i.e., as (+) or (−)-isomersrespectively). A chiral compound can exist as either individualenantiomer or as a mixture thereof. A mixture containing equalproportions of the enantiomers is called a “racemic mixture”.

“Tautomers” refer to compounds that are interchangeable forms of aparticular compound structure, and that vary in the displacement ofhydrogen atoms and electrons. Thus, two structures may be in equilibriumthrough the movement of π electrons and an atom (usually H). Forexample, enols and ketones are tautomers because they are rapidlyinterconverted by treatment with either acid or base. Another example oftautomerism is the aci- and nitro- forms of phenylnitromethane, that arelikewise formed by treatment with acid or base.

Tautomeric forms may be relevant to the attainment of the optimalchemical reactivity and biological activity of a compound of interest.

The compounds of this invention may possess one or more asymmetriccenters; such compounds can therefore be produced as individual (R)— or(S)— stereoisomers or as mixtures thereof. Unless indicated otherwise,the description or naming of a particular compound in the specificationand claims is intended to include both individual enantiomers andmixtures, racemic or otherwise, thereof. The methods for thedetermination of stereochemistry and the separation of stereoisomers arewell-known in the art.

THE COMPOUNDS

The present invention is based on the discovery of that MAPKAPK5functions in the pathway that results in the expression of MMP1, andthat inhibitors of MAPKAPK5 activity, such as the compounds of thepresent invention, are useful for the treatment of diseases involvingthe abnormally high expression of MMP activity.

The present matrix metallo proteinase inhibiting compounds of thepresent invention may be described generally asimidazo[1,2-a]pyrazine-8-yl-amines substituted in the 5-position by anaryl and heteroaryl group, and an in the 8-position by an arylamino or aheteroarylamino group.

More particularly, the present invention relates to compounds havingmatrix metallo proteinase inhibiting properties in a mammalian cell,according to formula (I):

wherein:

-   A and B are independently CR4R″, NR″, oxygen or sulfur;-   AA is CR4 or N;-   D is C═O, CR4R″ or NR″;-   E is NH or CR″R6, when k is zero, and is NH or CR″R6a, when k is    one;-   F is sulfur, oxygen or NH;-   T is oxygen or NR;-   U, V, W and X are independently CR″R7 or NR″;-   Y is CR″ or N;-   Z is hydrogen, amino, hydroxyl, lower alkoxy, carbamoyl, carboxyl,    SO₂Rz, SO₂NRRz, —NR(CO)(CH2)d-Rz, —NRRz, —(CO)—ORz,    —(CO)—NR(CH2)d-Rz, or

-   R is independently hydrogen or lower alkyl;-   R′ is independently hydrogen or lower alkyl;-   R″ is H or forms a double bond with an adjacent atom;-   R1 is H; R4; or lower alkyl, lower cycloalkyl and lower alkyl-lower    cycloalkyl, optionally substituted with one or more R4;-   R2 is H; lower alkyl, lower cycloalkyl and lower alkyl-lower    cycloalkyl, optionally substituted with one or more of F and Cl;-   R3 is H or forms a double bond with an adjacent R″;-   R4 is H, F, Cl; CN; COOR5; OR5; C(O)N(R5R5a); S(O)₂N(R5R5a); lower    alkyl; O— lower alkyl NH-lower alkyl; S-lower alkyl; COO— lower    alkyl; OC(O)— lower alkyl; C(O)N(R5)— lower alkyl; S(O)₂N(R5)-lower    alkyl; S(O)N(R5)-lower alkyl; S(O)₂-lower alkyl; S(O)-lower alkyl;    N(R5)S(O)₂-lower alkyl; and N(R5)S(O)-lower alkyl; wherein each    lower alkyl is optionally substituted with one or more of F and Cl;-   R5 and R5a are independently    -   H; F, Cl; or lower alkyl, lower cycloalkyl, or lower alkyl-lower        cycloalkyl optionally substituted with one or more of F and Cl;-   R6 is hydrogen, amino, hydroxyl, carbamoyl, carboxyl, SO₂R, NRR′,    —(CO)—OR, or —(CO)—NRR′;-   R6a is R6, Cl, F, lower alkoxy, cyano, trifluoromethoxy; or together    with the adjacent be —(CHR″)_(n)—NR—(CHR″)_(p)—, and form a five or    six member heterocyclic ring fused to the ring to which they are    bonded;-   R7 is independently hydrogen, halogen, lower alkyl or lower alkoxy;-   Rz is hydrogen, lower alkyl, lower alkanoyl, phenyl, 1-loweralkyl    pyrrolidin-3-yl, pyrazol-4-yl, pyrazol-2-yl, or lower alkyl, lower    alkanoyl, phenyl, 1-loweralkyl pyrrolidin-3-yl, pyrazol-4-yl,    pyrazol-2-yl or pyrid-3-yl substituted by one or more of hydroxyl,    amino, mono- or di- loweralkylamino, acetamidyl, lower alkanoyl,    lower alkyl, 4-hydroxy-phenyl, 3-aminomethylphenyl, lower alkyl    sulfonyl, 4-diloweralkylaminophenyl, pyrid-3-yl, 1H-indol-3-yl,    morpholin-4-yl,;-   R and Rz together may be —(CHR)_(q)-T-(CHR)_(r), and form a five or    six member heterocyclic ring with the nitrogen to which they are    bonded;-   Rz and R7 together may be —(CHR″)_(n)—NR—(CHR″)_(p)—, and form a    five or six member heterocyclic ring fused to the ring to which they    are bonded;-   b and d are independently 0 or 1; provided at least one of b or d is    1;-   k is 0 or 1;-   m is 0 or 1;-   n and p are independently 0, 1 or 2;-   q and r are 1 or 2;-   x is o or 1;-   with the provisos that: (1) when m is zero, and (a) either U or W is    NR″, then Z is not carbamoyl; and    -   (b) x is 1 and U is NR″, then W is not NR″;    -   (2) (a) when m is 1, or (b) when U, V, W, X and Y form phenyl        and X is C-lower alkoxy, or (c) when Rz together with R7 form        indolyl, then R6 is not carbamoyl; and    -   (3) at least one of :R7 is other than hydrogen;        or a pharmaceutically acceptable salt, hydrate, solvate or        prodrug thereof.

A preferred aspect of the present invention is a subclass of compoundsaccording to formula II,

wherein

-   A and B are independently CR4R″, NR″, oxygen or sulfur;-   AA is CR4 or N;-   D is C═O, CR4R″ or NR″;-   E is NH or CR″R6, when k is zero, and is NH or CR″R6a, when k is    one;-   F is sulfur, oxygen or NH;-   T is oxygen or NR;-   R is independently hydrogen or lower alkyl;-   R′ is independently hydrogen or lower alkyl;-   R″ is H or forms a double bond with an adjacent atom;-   R1 is H; R4; or lower alkyl, lower cycloalkyl and lower alkyl-lower    cycloalkyl, optionally substituted with one or more R4;-   R2 is H; lower alkyl, lower cycloalkyl and lower alkyl-lower    cycloalkyl, optionally substituted with one or more of F and Cl;-   R4 is H, F, Cl; CN; COOR5; OR5; C(O)N(R5R5a); S(O)₂N(R5R5a); lower    alkyl; O— lower alkyl; NH-lower alkyl; S-lower alkyl; COO— lower    alkyl; OC(O)— lower alkyl; C(O)N(R5)— lower alkyl; S(O)₂N(R5)-lower    alkyl; S(O)N(R5)-lower alkyl; S(O)₂-lower alkyl; S(O)-lower alkyl;    N(R5)S(O)₂-lower alkyl; and N(R5)S(O)-lower alkyl; wherein each    lower alkyl is optionally substituted with one or more of F and Cl;-   R5 and R5a are independently    -   H; F, Cl; or lower alkyl, lower cycloalkyl, or lower alkyl-lower        cycloalkyl optionally substituted with one or more of F and Cl;-   R6 is hydrogen, amino, hydroxyl, carbamoyl, carboxyl, SO₂R, NRR′,    —(CO)—OR, or —(CO)—NRR′;-   R6a is R6, Cl, F, lower alkoxy, cyano, trifluoromethoxy; or together    with the adjacent be —(CHR″)_(n)—NR—(CHR″)_(p)—, and form a five or    six member heterocyclic ring fused to the ring to which they are    bonded;-   R8 is phenyl independently substituted by R_(a) in the    ortho-position, by R_(b) in the meta-position, and by R_(c) in the    para-position; pyrid-3-yl; pyrid-3-yl substituted by R_(c) in the    5-position; or cyclohexyl independently substituted by R_(a) in the    2-position, and by R_(d) in the 4-position;    -   R_(a) is hydrogen, halogen, lower alkyl, trifluoromethyl or        lower alkoxy;    -   R_(b) is hydrogen, trifluoromethyl, lower alkoxy, lower alkyl        sulfonamide, carboxyl, —NR_(e)R_(f), —(CO)—OR or        —(CO)—NR_(e)R_(f);    -   R_(c) is hydrogen, amino, hydroxyl, lower alkoxy, carbamoyl,        carboxyl, SO₂R, SO₂N_(e)R_(f), NR_(e)R_(f), —(CO)—OR, or        —(CO)—NR_(e)R_(f); or    -   R_(b) and R_(c) can together form a benzdiazole, or indole        substituted in the 3-position by R′;    -   R_(d) is hydroxyl, halogen, amino, lower alkoxy, or NR_(e)R_(f);    -   R_(e) and R_(f) are independently hydrogen, 1-loweralkyl        pyrrolidin-3-yl, 1-R-pyrazol-4-yl, lower alkanoyl, phenyl, or        lower alkyl optionally substituted by one or more of        4-hydroxy-phenyl, 3-aminomethylphenyl, lower alkyl sulfonyl,        4-diloweralkylaminophenyl, pyrid-3-yl, 1H-indol-3-yl,        morpholin-4-yl, hydroxyl, amino, mono- or di-loweralkylamino, or        by lower alkanoyl; or R′ and R″ together are        —(CHR)_(n)-T-(CHR)_(n)— and form a five or six member        heterocyclic ring with the nitrogen to which they are bonded;-   m is 0, 1, or2;-   n is 1 or 2;    -   with the proviso that: (1) (a) when R8 is pyridy-3-yl, or (b)        when R_(b) is lower alkoxy or (c) together with        -   R_(c) are indolyl, then R_(e) is not carbamoyl;        -   (2) when R_(e) is hydroxyl, then R_(g) is not lower alkyl;            and        -   (3) at least one of R_(a), R_(b) and R_(c) is other than            hydrogen;            or a pharmaceutically acceptable salt, hydrate, solvate or            prodrug thereof.

Another aspect of the present invention relates to compounds accordingto formula III:

wherein

R¹ is H, or substituted or unsubstituted alkyl; R² is H, lower alkyl,lower cycloalkyl and lower alkyl-lower cycloalkyl, optionallysubstituted with one or more of F and Cl; R⁸ is selected fromsubstituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl, substituted orunsubstituted pyridyl, substituted or unsubstituted pyrimidine, andsubstituted or unsubstituted pyrazine, substituted or unsubstitutedpyrrole, substituted or unsubstituted pyrazole and substituted orunsubstituted imidazole; and R⁹ is selected from substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, substituted or unsubstitutedheteroaryl; or a pharmaceutically acceptable salt, solvate or prodrugthereof; and stereoisomers, isotopic variants and tautomers thereof.

In one embodiment, with respect to compounds of formula III, R¹ is H,Me, Et, i-Pr or CF₃.

In one embodiment, with respect to compounds of formula III, R′ is H.

In one embodiment, with respect to compounds of formula III, R² is H,Me, Et, i-Pr or CF₃.

In one embodiment, with respect to compounds of formula III, R² is H.

In one embodiment, with respect to compounds of formula III, R⁸ isselected from substituted or unsubstituted cycloalkyl.

In another embodiment, with respect to compounds of formula III, R⁸ isselected from substituted or unsubstituted cyclohexyl or cyclopentyl.

In one embodiment, with respect to compounds of formula III, R⁸ isselected from substituted or unsubstituted heterocycloalkyl.

In another embodiment, with respect to compounds of formula III, R⁸ isselected from substituted or unsubstituted piperidinyl, morpholinyl, orpyrrolidinyl.

In one embodiment, with respect to compounds of formula III, R⁸ isselected from substituted or unsubstituted phenyl, pyridyl orpyrimidine.

In one embodiment, with respect to compounds of formula III, R⁸ isselected from substituted phenyl, substituted pyridyl, and substitutedpyrimidine; and the substitution is

-   -   -L-R^(8d); and wherein    -   L is selected from a bond, alkylene, heteroalkylene, —O—,        —N(R^(8e))—, —CO—, —CO₂—, —SO—, —SO₂—, —CON(R^(8e))—,        —SO₂N(R^(8e))—, —N(R^(8e))CO—, —N(R^(8e))SO₂—, —N(R^(8e))CO        N(R^(8e))—, —N(R^(8e))SO₂N(R^(8e))—; and    -   R^(8d) is selected from substituted or unsubstituted alkyl,        substituted or unsubstituted cycloalkyl, substituted or        unsubstituted aryl, substituted or unsubstituted        heterocycloalkyl, substituted or unsubstituted heteroaryl,        substituted or unsubstituted amino, substituted or unsubstituted        aralkyl, substituted or unsubstituted heteroarylalkyl and        substituted or unsubstituted aminoalkyl; and    -   R^(8e) is selected from H, substituted or unsubstituted alkyl        and substituted or unsubstituted cycloalkyl.

In one embodiment, with respect to compounds of formula III, R⁸ is

wherein L, and R^(8d) are as described in the preceding paragraph; thesubscript n is selected from 1-4; and each R^(8a), is independentlyselected from hydrogen, substituted or unsubstituted alkyl, alkoxy,cyano, and halo.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above and the subscript n is 1 and R8a is Me, Et, Pr, iso-Pr,Cl, F, CN, OMe, or CF₃. In another embodiment, R⁸, is at 2-(ortho to -L)position. In yet another embodiment, R^(8a) is 2-Cl, 2-F, 2-Me or 2-CF₃.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above and

-   -   L is —CON(R^(8e))— or SO₂N(R^(8e)e)—;    -   R^(8d) is selected from substituted or unsubstituted alkyl,        substituted or unsubstituted cycloalkyl, substituted or        unsubstituted aryl, substituted or unsubstituted        heterocycloalkyl, substituted or unsubstituted heteroaryl,        substituted or unsubstituted aralkyl, substituted or        unsubstituted heteroarylalkyl and substituted or unsubstituted        aminoalkyl; and    -   R^(8e) is selected from H, substituted or unsubstituted alkyl.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above and

-   -   L is —CONH— or SO₂NH—; and    -   R^(8d) is selected from H, alkylaminoethyl, dialkylaminoethyl,        cycloalkyl, heterocycloalkyl, arylalkyl, and heteroarylalkyl.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above and

-   -   L is —CONH— or SO₂NH—; and    -   R^(8d) is selected from methylaminoethyl, ethylaminoethyl,        dimethylaminoethyl, diethylaminoethyl, substituted or        unsubstituted pyrrolidinyl, benzyl and pyridylmethyl.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above and

-   -   L is a bond, —CO—, SO₂, —(CH₂)_(m1)—, —O(CH₂)_(m1)—,        —NH(CH₂)_(m1)—,—CON(H)(CH₂)_(m1)—, or —SO₂NH(CH₂)_(m1)—; the        subscript m1 is selected from 1-4; and R^(8d) is

-   -    and wherein the ring P is substituted or unsubstituted        heterocycloalkyl.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above; L is a bond; and the ring P is substituted orunsubstituted heterocycloalkyl.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above; L is a bond; and the ring P is substituted orunsubstituted piperidine, substituted or unsubstituted piperazine, andsubstituted or unsubstituted piperidine, morpholine.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above; L is CO or SO₂; and the ring P is substituted orunsubstituted heterocycloalkyl.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above; L is CO or SO₂; and the ring P is substituted orunsubstituted piperidine, substituted or unsubstituted piperazine, andsubstituted or unsubstituted piperidine, morpholine.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above; L is —(CH₂)_(m1)—, —O(CH₂)_(m1)—, or —NH(CH₂)_(m1)-;the subscript m1 is selected from 1-4; and the ring P is substituted orunsubstituted heterocycloalkyl.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above; L is —(CH₂)_(m1)—, —O(CH₂)_(m1)—, or —NH(CH₂)_(m1)—;the subscript m1 is 2 or 3; and the ring P is substituted orunsubstituted piperidine, substituted or unsubstituted piperazine, andsubstituted or unsubstituted piperidine, morpholine.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above; L is —CON(H)(CH₂)_(m1)—, or —SO₂NH(CH₂)_(m1)—; thesubscript m1 is selected from 1-4; and the ring P is substituted orunsubstituted heterocycloalkyl.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above; L is —CON(H)(CH₂)_(m1)—, or —SO₂NH(CH₂)_(m1)—; thesubscript ml is 2 or 3; and the ring P is substituted or unsubstitutedpiperidine, substituted or unsubstituted piperazine, and substituted orunsubstituted piperidine, morpholine.

In one embodiment, with respect to compounds of formula III, thecompound is according to formula IVa, IVb, IVc, or IVd:

and wherein L is a bond, —CO—, SO₂, —(CH₂)_(m1)—, —O(CH₂)_(m1)—,—NH(CH₂)_(m1)—, —CON(H)(CH₂)_(m1)—, or —SO₂NH(CH₂)_(m1)—; the subscriptm1 is selected from 1-4; the ring P is substituted or unsubstitutedheterocycloalkyl; the subscript n is selected from 1-4; each R^(8a) isindependently selected from hydrogen, substituted or unsubstitutedalkyl, alkoxy, cyano, and halo; and R⁹ is independently selected fromsubstituted or unsubstituted aryl and heteroaryl; or a pharmaceuticallyacceptable salt, solvate or prodrug thereof; and stereoisomers, isotopicvariants and tautomers thereof.

In one embodiment, with respect to compounds of formulae IVa-IVd, L is abond.

In one embodiment, with respect to compounds of formulae IVa-IVd, L ismethylene, ethylene, propylene, and butylene.

In one embodiment, with respect to compounds of formulae IVa-IVd, L is—CO—.

In one embodiment, with respect to compounds of formulae IVa-IVd, L is—SO₂—.

In one embodiment, with respect to compounds of formulae IVa-IVd, L is—CON(H)—CH₂—CH₂—, or —SO₂NH—CH₂—CH₂—.

In one embodiment, with respect to compounds of formulae IVa-IVd, L is—OCH₂—CH₂— or —NHCH₂—CH₂—.

In a preferred embodiment L is a bond, CO or SO₂.

In one embodiment, with respect to compounds of formulae IVa-IVd, thering P is substituted or unsubstituted piperidine, morpholine orpiperazine.

In one embodiment, with respect to compounds of formulae IVa-IVd, L andthe ring P are as described above; the subscript n is 4 and each R^(8a)is H.

In one embodiment, with respect to compounds of formulae IVa-IVd, L andthe ring P are as described above; the subscript n is 1 and R^(8a) isMe, Et, Pr, iso-Pr, Cl, F, CN, OMe, or CF₃. In another embodiment,R^(8a) is at 2-(ortho to -L) position. In yet another embodiment, R^(8a)is 2-Cl, 2-F, 2-Me or 2-CF₃.

In a further embodiment, with respect to compounds of formula III, R⁸ is

and wherein the ring P is substituted or unsubstituted heterocycloalkyl;the subscript n is selected from 1-4 and each R^(8a) is independentlyselected from hydrogen, substituted or unsubstituted alkyl, alkoxy,cyano, and halo.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above and the ring P is substituted or unsubstitutedpiperidine, morpholine or piperazine.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above and the subscript n is 4 and each R^(8a) is H.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above and the subscript n is 1 and R^(8a) is Me, Et, Pr,iso-Pr, Cl, F, CN, OMe, or CF₃. In another embodiment, R^(8a) is at the2-(ortho to N-ring P) position. In yet another embodiment, R^(8a) is2-Cl, 2-F, 2-Me or 2-CF₃.

In a further embodiment, with respect to compounds of formula III, R⁸ is

and wherein the subscript n is selected from 1-4; each R^(8a) isindependently selected from hydrogen, substituted or unsubstitutedalkyl, alkoxy, cyano, and halo; R^(8b) is hydrogen, substituted orunsubstituted alkyl or substituted or unsubstituted cycloalkyl; R^(8c)is hydrogen, substituted or unsubstituted alkyl and the subscript x isselected from 1-8.

In a further embodiment, with respect to compounds of formula III, R⁸ is

and wherein the subscript n is selected from 1-4; each R^(8a) isindependently selected from hydrogen, substituted or unsubstitutedalkyl, alkoxy, cyano, and halo; R^(8b) is hydrogen, substituted orunsubstituted alkyl or substituted or unsubstituted cycloalkyl; R^(8c)is hydrogen or Me.

In a further embodiment, with respect to compounds of formula III, R⁸ is

and wherein the subscript n is selected from 1-4; each R^(8a) isindependently selected from hydrogen, substituted or unsubstitutedalkyl, alkoxy, cyano, and halo.

In a further embodiment, with respect to compounds of formula III, Ru⁸is

and wherein the subscript n is selected from 1-4; each R^(8a) isindependently selected from hydrogen, substituted or unsubstitutedalkyl, alkoxy, cyano, and halo; and R^(8b) is hydrogen, substituted orunsubstituted alkyl or substituted or unsubstituted cycloalkyl.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above and the subscript n is 4 and each R^(8a) is H.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above and the subscript n is 1 and R^(8a) is Me, Et, Pr,iso-Pr, Cl, F, CN, OMe, or CF₃. In another embodiment, R^(8a) is at2-(ortho to N-ringP) position. In yet another embodiment, R^(8a) is2-Cl, 2-F, 2-Me or 2-CF₃.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above and R^(8b) is H.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above and R^(8b) substituted or unsubstituted alkyl.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above and R^(8b) substituted or unsubstituted cycloalkyl.

In one embodiment, with respect to compounds of formula III, R⁸ is asdescribed above and R^(8b) is Me, Et, Pr, i-Pr, t-Bu, i-Bu, CH₂CF₃, CF₃,CH₂CONH₂, cyclopropyl or cyclopropylmethyl.

In one particular embodiment, with respect to compounds of formula III,R⁸ is as described above and R^(8b) is i-Pr.

In a further embodiment, with respect to compounds of formulae IVa-IVd,R^(8a) is independently selected from hydrogen, substituted orunsubstituted alkyl, alkoxy, cyano, carbamoyl, CHO, and halo. In oneembodiment, R^(8a) is H, Me, F, or Cl. In a preferred embodiment R^(8a)is H.

In one embodiment, with respect to compounds of formulae IVa-IVd, thecompound is according to formula Va, Vb, Vc, Vd, Ve, or Vf:

and wherein R⁹ is as described for formula III and R^(8b) is hydrogen,substituted or unsubstituted alkyl or substituted or unsubstitutedcycloalkyl.

In one embodiment, with respect to compounds of formulae Va-Vf, R^(8b)is H.

In one embodiment, with respect to compounds of formulae Va-Vf, R^(8b)is substituted or unsubstituted alkyl.

In one embodiment, with respect to compounds of formulae Va-Vf, R^(8b)is substituted or unsubstituted cycloalkyl.

In one embodiment, with respect to compounds of formulae Va-Vf, R^(8b)is Me, Et, Pr, i-Pr, t-Bu, i-Bu, CH₂CF₃, CF₃, CH₂CONH₂, cyclopropyl orcyclopropylmethyl.

In one particular embodiment, with respect to compounds of formulaeVa-Vf, R^(8b) is i-Pr.

In a one embodiment, with respect to compounds of formulae III-VF, R⁹ issubstituted or unsubstituted aryl. In another embodiment, R⁹ issubstituted or unsubstituted phenyl.

In a one embodiment, with respect to compounds of formulae III-VF, R⁹ issubstituted or unsubstituted heteroaryl. In another embodiment, R⁹ issubstituted or unsubstituted pyridyl.

In a one embodiment, with respect to compounds of formulae III-VF, R⁹ isselected from substituted or unsubstituted phenyl, indolyl, isoinolyl,pyrrolyl, furanyl, thienyl, pyrazolyl, oxazolyl, and thiazolyl.

In one embodiment, with respect to compounds of formulae III-Vf, R⁹ is

and each of A¹, A² and A³ is independently selected from S, O, N,NR^(9a), and CR^(9a); each of R^(9a) is independently H or substitutedor unsubstituted alkyl; and R^(9b) is CONH₂, CONHMe, or CN.

In further embodiment, with respect to compounds of formulae III-Vf, R⁹is

In further embodiment, with respect to compounds of formulae III-Vf, R⁹is

In further embodiment, with respect to compounds of formulae III-Vf, R⁹is

and wherein the subscript m is selected from 1-4 and each R^(9d) isindependently H, substituted or unsubstituted alkyl or halo.

In further embodiment, with respect to compounds of formulae III-Vf, R⁹is

and wherein the subscript m is selected from 1-4 and each R^(9d) isindependently H, substituted or unsubstituted alkyl or halo.

In a further embodiment, with respect to compounds of formulae III-VF,R⁹ is

and wherein the subscript m is selected from 1-3 and each R^(9d) isindependently H, substituted or unsubstituted alkyl or halo.

In a further embodiment, with respect to compounds of formulae III-VF,R⁹ is as described above; and each R^(9d) is H.

In a further embodiment, with respect to compounds of formulae III-VF,R⁹ is as described above; m is 1 or 2 and each R^(9d) is independentlyMe, Cl or F.

In one embodiment, with respect to compounds of formula III, thecompound is according to formula VIa, VIb, Vic, VId, VIe or VIf:

and R^(8b) is hydrogen, substituted or unsubstituted alkyl orsubstituted or unsubstituted cycloalkyl.

In a further embodiment, with respect to compounds of formulae VIa-VIf,R^(8b) is H.

In a further embodiment, with respect to compounds of formulae VIa-VIf,R^(8b) is cycloalkyl.

In a further embodiment, with respect to compounds of formulae VIa-VIf,R^(8b) is cyclopropyl.

In a further embodiment, with respect to compounds of formulae VIa-VIf,R^(8b) is substituted or unsubstituted alkyl.

In a further embodiment, with respect to compounds of formulae VIa-VIf,R^(8b) is Me, Et, Pr, i-Pr, t-Bu, i-Bu, CF₃, CH₂CF₃, CH₂CONH₂, orcyclopropylmethyl.

In one embodiment, with respect to compounds of formula III, thecompound is according to formula VIIa, VIb, VIIc, VIIc, VId, VIIe orVIIf:

and R^(8b) is hydrogen, substituted or unsubstituted alkyl orsubstituted or unsubstituted cycloalkyl.

In a further embodiment, with respect to compounds of formulaeVIIa-VIId, R^(8b) is H.

In a further embodiment, with respect to compounds of formulaeVIIa-VIIIf, R⁸b is cycloalkyl.

In a further embodiment, with respect to compounds of formulaeVIIa-VIIf, R^(8b) is cyclopropyl.

In a further embodiment, with respect to compounds of formulaeVIIa-VIIf, R^(8b) is substituted or unsubstituted alkyl.

In a further embodiment, with respect to compounds of formulae VIa-VIIf,R^(8b) is Me, Et, Pr, i-Pr, t-Bu, i-Bu, CF₃, CH₂CF₃, CH₂CONH₂, orcyclopropylmethyl.

In one embodiment, with respect to compounds of formula III, thecompound is according to formula VIIa, VIIb, VIIc, VIId, VIIe or VIIIf:

and R^(8b) is hydrogen, substituted or unsubstituted alkyl orsubstituted or unsubstituted cycloalkyl.

In a further embodiment, with respect to compounds of formulaeVIIIa-VIIIf, R^(8b) is H.

In a further embodiment, with respect to compounds of formulaeVIIIa-VIIIf, R^(8b) is cycloalkyl.

In a further embodiment, with respect to compounds of formulaeVIIIa-VIIIf, R^(8b) is cyclopropyl.

In a further embodiment, with respect to compounds of formulaeVIIIa-VIIIf, R^(8b) is substituted or unsubstituted alkyl.

In a further embodiment, with respect to compounds of formulaeVIIIa-VIIIf, R^(8b) is Me, Et, Pr, i-Pr, t-Bu, i-Bu, CF₃, CH₂CF₃,CH₂CONH₂, or cyclopropylmethyl.

In one embodiment, with respect to compounds of formula III, thecompound is according to formula IXa, IXb, IXc, IXd, IXe, or IXf:

and R^(8b) is hydrogen, substituted or unsubstituted alkyl orsubstituted or unsubstituted cycloalkyl.

In a further embodiment, with respect to compounds of formulae IXa-IXf,R^(8b) is H.

In a further embodiment, with respect to compounds of formulae IXa-IXf,R^(8b) is cycloalkyl.

In a further embodiment, with respect to compounds of formulae Ixa-IXf,R^(8b) is cyclopropyl.

In a further embodiment, with respect to compounds of formulae IXa-IXf,R^(8b) is substituted or unsubstituted alkyl.

In a further embodiment, with respect to compounds of formulae IXa-IXf,R^(8b) is Me, Et, Pr, i-Pr, t-Bu, i-Bu, CF₃, CH₂CF₃, CH₂CONH₂, orcyclopropylmethyl.

In one embodiment, with respect to compounds of formula III, thecompound is according to formula Xa, Xb, Xc, Xd, Xe, or Xf:

and R^(8b) is hydrogen, substituted or unsubstituted alkyl orsubstituted or unsubstituted cycloalkyl.

In a further embodiment, with respect to compounds of formulae Xa-Xf,R^(8b) is H.

In a further embodiment,.with respect to compounds of formulae Xa-Xf,R^(8b) is cycloalkyl.

In a further embodiment, with respect to compounds of formulae Xa-Xf,R^(8b) is cyclopropyl.

In a further embodiment, with respect to compounds of formulae Xa-Xf,R^(8b) is substituted or unsubstituted alkyl.

In a further embodiment, with respect to compounds of formulae Xa-Xf,R^(8b) is Me, Et, Pr, i-Pr, t-Bu, i-Bu, CF₃, CH₂CF₃, CH₂CONH₂, orcyclopropylmethyl.

In one embodiment, with respect to compounds of formula III, thecompound is according to formula XIa, XIb, XIc, XId, XIe or XIf:

and R^(8b) is hydrogen, substituted or unsubstituted alkyl orsubstituted or unsubstituted cycloalkyl; and R^(9e) is hydrogen, Me, orCN.

In one embodiment, with respect to compounds of formulae XIa-XIf, R^(9e)is H.

In one embodiment, with respect to compounds of formulae XIa-XIf, R^(9e)is Me.

In one embodiment, with respect to compounds of formulae XIa-XIf, R^(9e)is CN.

In a further embodiment, with respect to compounds of formulae XIa-XIf,R^(8b) is H.

In a further embodiment, with respect to compounds of formulae XIa-XIf,R^(8b) is cycloalkyl.

In a further embodiment, with respect to compounds of formulae XIa-XIf,R^(8b) is cyclopropyl.

In a further embodiment, with respect to compounds of formulae XIa-XIf,R^(8b) is substituted or unsubstituted alkyl.

In a further embodiment, with respect to compounds of formulae XIa-XIf,R^(8b) is Me, Et, pr, i-Pr, t-Bu, i-Bu, CF₃, CH₂CF₃, CH₂CONH₂, orcyclopropylmethyl.

In one embodiment, with respect to compounds of formula III, thecompound is according to formula XIIa, XIIb, XIIc or XIId:

In one embodiment, with respect to compounds of formula III, thecompound is according to formula XIIIa, XIIIb, XIIIc or XIIId:

In one embodiment, with respect to compounds of formula III, thecompound is according to formula XIVa, XIVb, XIVc or XIVd:

In one embodiment, with respect to compounds of formula III, thecompound is according to formula XVa, XVb, or XIVc:

-   -   and L is a bond, —CO—, SO₂, —(CH₂)_(m1)—, —O(CH₂)_(m1)—,        —NH(CH₂)_(m1)—, —CON(H)(CH₂)_(m1)—, or —SO₂NH(CH₂)_(m1)—; the        subscript m1 is selected from 1-4; the ring P is

and R^(8b) is H, Me, i-Pr, t-Bu, CH₂CONH₂, cyclopropylmethyl, or CH₂CF₃.

In one particular embodiment, with respect to compounds of formulaeXVa-XVc, L is a bond. In another particular embodiment, L is—O—CH₂—CH₂—.

In one particular embodiment, with respect to compounds of formulaeXVa-XVc, the ring P is

In more particular embodiment, with respect to compounds of formulaeXVa-XVc, the ring P is

In another embodiment, with respect to compounds of formula III, thecompound is selected from Table 1.

In another embodiment, with respect to compounds of formula III, thecompound is selected from:

-   -   4-(8-{4-[Ethyl-(2-hydroxy-ethyl)-amino]-phenylamino}-imidazo[1,2-a]pyrazin-5-yl)-3-methyl-phenol;    -   4-(8-{4-[Ethyl-(2-hydroxy-ethyl)-amino]-phenylamino}-imidazo[1,2-a]pyrazin-5-yl)-benzamide;    -   4-(8-{4-[Ethyl-(2-hydroxy-ethyl)-amino]-phenylamino}-imidazo[1,2-a]pyrazin-5-yl)-phenol;    -   2-({4-[5-(4-Amino-phenyl)-imidazo[1,2-a]pyrazin-8-ylamino]-phenyl}-ethyl-amino)-ethanol;    -   2-({4-[5-(3-Amino-phenyl)-imidazo[1,2-a]pyrazin-8-ylamino]-phenyl}-ethyl-amino)-ethanol;    -   3-[5-(1H-Pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide;    -   2-(Ethyl-{3-methyl-4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-phenyl}-amino)-ethanol;    -   N-[5-(1H-Pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-benzene-1,4-diamine;    -   4-[3-Methyl-8-(4-morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-benzamide;    -   [3-Methyl-5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-(4-morpholin-4-yl-phenyl)-amine;    -   4-[3-Methyl-8-(4-morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-1H-pyridin-2-one;    -   4-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-6-trifluoromethyl-1H-pyridin-2-one;    -   4-[3-Ethyl-8-(4-morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-benzamide;    -   6-Methyl-4-[8-(4-morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-pyridin-2-ol;    -   4-[3-Ethyl-8-(4-morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-1H-pyridin-2-one;    -   [3-Ethyl-5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-(4-morpholin-4-yl-phenyl)-amine;    -   4-(8-{4-[1-(2,2,2-Trifluoro-ethyl)-piperidin-4-yl]-phenylamino}-imidazo[1,2a]pyrazin-5-yl)-thiophene-2-carboxylic        acid amide;    -   [5-(1H-Pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-{4-[1-(2,2,2-trifluoro-ethyl)-piperidin-4-yl]-phenyl}-amine;    -   4-{8-[4-(1-Isopropyl-piperidin-4-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-thiophene-2-carboxylic        acid amide;    -   [6-(4-Isopropyl-piperazin-1-yl)-pyridin-3-yl]-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine;    -   [4-(1-Isopropyl-piperidin-4-yl)-phenyl]-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine;    -   5-{8-[4-(1-Isopropyl-piperidin-4-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-2,3,        -dihydro-isoindol-1-one;    -   5-[8-(6-Morpholin-4-yl-pyridin-3-ylamino)-imidazo[1,2-a]pyrazin-5-yl]-2,3-dihydo-isoindol-1-one;    -   5-[8-(2-Morpholin-4-yl-pyrimidin-5-ylamino)-imidazo[1,2-a]pyrazin-5-yl]-2,3-dihydro-isoindol-1-one;    -   2,6-Difluoro-4-{8-[4-(2-morpholin-4-yl-ethoxy)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide;    -   2-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-5,6-dihydro-furo[2,3-c]pyrrol-4-one;    -   (3-Dimethylaminomethyl-4-morpholin-4-yl-phenyl)-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine;    -   5-{8-[2-(4-Isopropyl-piperazin-1-yl)-pyrimidin-5-ylamino]-imidazo[1,2-a]pyrazin-5-yl}-2,3-dihydro-isoindol-1-one;    -   7-Fluoro-5-{8-[4-(4-isopropyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-2,3-dihydro-isoindol-1-one;    -   2-{8-[4-(4-Isopropyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-5,6-dihydro-furo[2,3-c]pyrrol-4-one;    -   2-(4-{4-[5-(1H-Pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-phenyl}-piperidin-1yl)-acetamide;    -   2-(4-{4-[5-(5-Methyl-1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-phenyl}-piperidin-1-yl)-acetamide;    -   2-(4-{4-[5-(1-Oxo-2,3-dihydro-1H-isoindol-5-yl)-imidazo[1,2-a]pyrazin-8ylamino]-phenyl}-piperidin-1-yl)-acetamide;    -   5-{8-[4-(4-Isopropyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-3,3-dimethyl-2,3-dihydro-isoindol-1-one;        and    -   2-{8-[4-(4-Isopropyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-4,5-dihydro-thieno[2,3-c]pyrrol-6-one.

In another embodiment, with respect to compounds of formula III, thecompound is selected from:

-   -   (4-Morpholin-4-yl-phenyl)-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine;    -   N,N-Diethyl-N′-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-benzene-1,4-diamine;    -   4-[5-(1H-Pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide;    -   (1H-Indol-5-yl)-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine;    -   N-{4-[5-(1H-Pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-phenyl}-methanesulfonamide;    -   N-{4-[5-(1H-Pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-phenyl}-acetamide;    -   N-(2-Diethylamino-ethyl)-4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide;    -   4-[5-(1H-Pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-N-pyridin-3-ylmethyl-benzamide;    -   4-[5-(1H-Pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzenesulfonamide;    -   Morpholin-4-yl-{4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-phenyl}-methanone;    -   N-(2-Morpholin-4-yl-ethyl)-4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide;    -   4-[5-(1H-Pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-N-pyrrolidin-3-yl-benzamide;    -   N-(2-Ethylamino-ethyl)-4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide;    -   N-(2-Hydroxy-ethyl)-4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide;    -   N-(1-Methyl-pyrrolidin-3-yl)-4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide;    -   N-(2-Diethylamino-ethyl)-4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzenesulfonamide;    -   [4-(4-Methyl-piperazine-1-sulfonyl)-phenyl]-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine;    -   [4-(4-Methyl-piperazine-1-sulfonyl)-phenyl]-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine;    -   N-(2-Diethylamino-ethyl)-2-fluoro-4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide;    -   N-(2-Diethylamino-ethyl)-3-fluoro-4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide;    -   N-(2-Diethylamino-ethyl)-4-[5-(2H-[1,2,3]triazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide;    -   4-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-1H-pyridin-2-one;    -   N-(4-Hydroxy-benzyl)-4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide;    -   4-[5-(1H-Pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-N-(2-pyridin-3-yl-ethyl)-benzamide;    -   N-[2-(1H-Indol-3-yl)-ethyl]-4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide;    -   N-(2-Diethylamino-ethyl)-4-[5-(2-oxo-1,2-dihydro-pyridin-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide;    -   4-{8-[4-(4-Methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-1H-pyridin-2-one;    -   5-[5-(1H-Pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-pyridine-2-carboxylic        acid (2-diethylamino-ethyl)-amide;    -   4-[5-(2-Oxo-1,2-dihydro-pyridin-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-N-pyridin-3-ylmethyl-benzamide;    -   N-(4-Hydroxy-benzyl)-4-[5-(2-oxo-1,2-dihydro-pyridin-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide;    -   N-(2-Dimethylamino-2-pyridin-3-yl-ethyl)-4-[5-(2-oxo-1,2-dihydro-pyridin-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide;    -   4-{8-[4-(4-Methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-thiophene-2-carboxylic        acid amide;    -   [4-(4-Methyl-piperazin-1-yl)-phenyl]-[5-(5-methyl-1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine;    -   2-Fluoro-4-{8-[4-(4-methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide;    -   3-Fluoro-4-{8-[4-(4-methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5yl}-benzamide;    -   5-{8-[4-(4-Methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-2-carboxylic        acid amide; and    -   5-{8-[4-(4-Isopropyl-piperazin-1-yl)-3-trifluoromethyl-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-2,3-dihydro-isoindol-1-one.

In certain aspects, the present invention provides prodrugs andderivatives of the compounds according to the formulae above. Prodrugsare derivatives of the compounds of the invention, which havemetabolically cleavable groups and become by solvolysis or underphysiological conditions the compounds of the invention, which arepharmaceutically active, in vivo. A prodrug may be inactive whenadministered to a subject but is converted in vivo to an active compoundof the invention. “Pharmaceutically acceptable prodrugs” as used hereinrefers to those prodrugs of the compounds useful in the presentinvention, which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of patients with unduetoxicity, irritation, allergic response commensurate with a reasonablebenefit/risk ratio, and effective for their intended use of thecompounds of the invention. The term “prodrug” means a compound that istransformed in vivo to yield an effective compound useful in the presentinvention or a pharmaceutically acceptable salt, hydrate or solvatethereof. The transformation may occur by various mechanisms, such asthrough hydrolysis in blood. The compounds bearing metabolicallycleavable groups have the advantage that they may exhibit improvedbioavailability as a result of enhanced solubility and/or rate ofabsorption conferred upon the parent compound by virtue of the presenceof the metabolically cleavable group, thus, such compounds act aspro-drugs. A thorough discussion is provided in Design of Prodrugs, H.Bundgaard, ed., Elsevier (1985); Methods in Enzymology; K. Widder et al,Ed., Academic Press, 42, 309-396 (1985); A Textbook of Drug Design andDevelopment, Krogsgaard-Larsen and H. Bandaged, ed., Chapter 5; “Designand Applications of Prodrugs” 113-191 (1991); Advanced Drug DeliveryReviews, H. Bundgard, 8, 1-38, (1992); J. Pharm. Sci., 77,285 (1988);Chem. Pharm. Bull., N. Nakeya et al, 32, 692 (1984); Pro-drugs as NovelDelivery Systems, T. Higuchi and V. Stella, 14 A.C.S. Symposium Series,and Bioreversible Carriers in Drug Design, E. B. Roche, ed., AmericanPharmaceutical Association and Pergamon Press, 1987, which areincorporated herein by reference. Such examples include, but are notlimited to, choline ester derivatives and the like, N-alkylmorpholineesters and the like.

Other derivatives of the compounds of this invention have activity inboth their acid and acid derivative forms, but the acid sensitive formoften offers advantages of solubility, tissue compatibility, or delayedrelease in the mammalian organism (see, Bundgard, H., Design ofProdrugs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985). Prodrugs includeacid derivatives well know to practitioners of the art, such as, forexample, esters prepared by reaction of the parent acid with a suitablealcohol, or amides prepared by reaction of the parent acid compound witha substituted or unsubstituted amine, or acid anhydrides, or mixedanhydrides. Simple aliphatic or aromatic esters, amides and anhydridesderived from acidic groups pendant on the compounds of this inventionare preferred prodrugs. In some cases it is desirable to prepare doubleester type prodrugs such as (acyloxy)alkyl esters or((alkoxycarbonyl)oxy)alkylesters. Preferred are the C₁ to C₈ alkyl,C₂-C₈ alkenyl, aryl, C₇-C₁₂ substituted aryl, and C₇-C₁₂ arylalkylesters of the compounds of the invention.

PHARMACEUTICAL COMPOSITIONS

When employed as pharmaceuticals, the compounds of this invention aretypically administered in the form of a pharmaceutical composition. Suchcompositions can be prepared in a manner well known in thepharmaceutical art and comprise at least one active compound.

Generally, the compounds of this invention are administered in apharmaceutically effective amount. The amount of the compound actuallyadministered will typically be determined by a physician, in the lightof the relevant circumstances, including the condition to be treated,the chosen route of administration, the actual compound -administered,the age, weight, and response of the individual patient, the severity ofthe patient's symptoms, and the like.

The pharmaceutical compositions of this invention can be administered bya variety of routes including oral, rectal, transdermal, subcutaneous,intravenous, intramuscular, and intranasal. Depending on the intendedroute of delivery, the compounds of this invention are preferablyformulated as either injectable or oral compositions or as salves, aslotions or as patches all for transdermal administration.

The compositions for oral administration can take the form of bulkliquid solutions or suspensions, or bulk powders. More commonly,however, the compositions are presented in unit dosage forms tofacilitate accurate dosing. The term “unit dosage forms” refers tophysically discrete units suitable as unitary dosages for human subjectsand other mammals, each unit containing a predetermined quantity ofactive material calculated to produce the desired therapeutic effect, inassociation with a suitable pharmaceutical excipient. Typical unitdosage forms include prefilled, premeasured ampules or syringes of theliquid compositions or pills, tablets, capsules or the like in the caseof solid compositions. In such compositions, the furansulfonic acidcompound is usually a minor component (from about 0.1 to about 50% byweight or preferably from about 1 to about 40% by weight) with theremainder being various vehicles or carriers and processing aids helpfulfor forming the desired dosing form.

Liquid forms suitable for oral administration may include a suitableaqueous or nonaqueous vehicle with buffers, suspending and dispensingagents, colorants, flavors and the like. Solid forms may include, forexample, any of the following ingredients, or compounds of a similarnature: a binder such as microcrystalline cellulose, gum tragacanth orgelatin; an excipient such as starch or lactose, a disintegrating agentsuch as alginic acid, Primogel, or corn starch; a lubricant such asmagnesium stearate; a glidant such as colloidal silicon dioxide; asweetening agent such as sucrose or saccharin; or a flavoring agent suchas peppermint, methyl salicylate, or orange flavoring.

Injectable compositions are typically based upon injectable sterilesaline or phosphate-buffered saline or other injectable carriers knownin the art. As before, the active compound in such compositions istypically a minor component, often being from about 0.05 to 10% byweight with the remainder being the injectable carrier and the like.

Transdermal compositions are typically formulated as a topical ointmentor cream containing the active ingredient(s), generally in an amountranging from about 0.01 to about 20% by weight, preferably from about0.1 to about 20% by weight, preferably from about 0.1 to about 10% byweight, and more preferably from about 0.5 to about 15% by weight. Whenformulated as a ointment, the active ingredients will typically becombined with either a paraffinic or a water-miscible ointment base.Alternatively, the active ingredients may be formulated in a cream with,for example an oil-in-water cream base. Such transdermal formulationsare well-known in the art and generally include additional ingredientsto enhance the dermal penetration of stability of the active ingredientsor the formulation. All such known transdermal formulations andingredients are included within the scope of this invention.

The compounds of this invention can also be administered by atransdermal device. Accordingly, transdermal administration can beaccomplished using a patch either of the reservoir or porous membranetype, or of a solid matrix variety.

The above-described components for orally administrable, injectable ortopically administrable compositions are merely representative. Othermaterials as well as processing techniques and the like are set forth inPart 8 of Remington's Pharmaceutical Sciences, 17th edition, 1985, MackPublishing Company, Easton, Pa., which is incorporated herein byreference.

The compounds of this invention can also be administered in sustainedrelease forms or from sustained release drug delivery systems. Adescription of representative sustained release materials can be foundin Remington's Pharmaceutical Sciences.

The following formulation examples illustrate representativepharmaceutical compositions of this invention. The present invention,however, is not limited to the following pharmaceutical compositions.

Formulation 1—Tablets

A compound of the invention is admixed as a dry powder with a drygelatin binder in an approximate 1:2 weight ratio. A minor amount ofmagnesium stearate is added as a lubricant. The mixture is formed into240-270 mg tablets (80-90 mg of active amide compound per tablet) in atablet press.

Formulation 2—Capsules

A compound of the invention is admixed as a dry powder with a starchdiluent in an approximate 1:1 weight ratio. The mixture is filled into250 mg capsules (125 mg of active amide compound per capsule).

Formulation 3—Liquid

A compound of the invention (125 mg), sucrose (1.75 g) and xanthan gum(4 mg) are blended, passed through a No. 10 mesh U.S. sieve, and thenmixed with a previously made solution of microcrystalline cellulose andsodium carboxymethyl cellulose (11:89, 50 mg) in water. Sodium benzoate(10 mg), flavor, and color are diluted with water and added withstirring. Sufficient water is then added to produce a total volume of 5mL.

Formulation 4—Tablets

A compound of the invention is admixed as a dry powder with a drygelatin binder in an approximate 1:2 weight ratio. A minor amount ofmagnesium stearate is added as a lubricant. The mixture is formed into450-900 mg tablets (150-300 mg of active amide compound) in a tabletpress.

Formulation 5—Injection

A compound of the invention is dissolved or suspended in a bufferedsterile saline injectable aqueous medium to a concentration ofapproximately 5 mg/ml.

Formulation 6—Topical

Stearyl alcohol (250 g) and a white petrolatum (250 g) are melted atabout 75° C. and then a mixture of a compound of the invention (50 g)methylparaben (0.25 g), propylparaben (0.15 g), sodium lauryl sulfate(10 g), and propylene glycol (120 g) dissolved in water (about 370 g) isadded and the resulting mixture is stirred until it congeals.

METHODS OF TREATMENT

The present compounds are used as therapeutic agents for the treatmentof conditions in mammals that are causally related or attributable toaberrant activity of MMP1 and/or MAPKAPK5. Accordingly, the compoundsand pharmaceutical compositions of this invention find use astherapeutics for preventing and/or treating inflammatory diseases inmammals including humans.

In a method of treatment aspect, this invention provides a method oftreating a mammal susceptible to or afflicted with a conditionassociated with extra-cellular matrix (ECM) degradation, in particulararthritis, and more particularly, rheumatoid arthritis which methodcomprises administering an effective amount of one or more of thecompounds of the invention or a pharmaceutical composition such as justdescribed.

In another method of treatment aspect, the invention provides a methodof treating a mammal susceptible to or afflicted with a conditionassociated with an abnormal cellular expression of MMP1, which comprisesadministering a therapeutically effective amount of a compound of theinvention, or a pharmaceutical composition thereof.

In another method of treatment aspect, the present invention provides amethod of treatment or prophylaxis of a condition characterized byabnormal matrix metallo proteinase activity, which comprisesadministering a therapeutically effective matrix metallo proteinaseinhibiting amount of one or more of the compounds of the invention, orpharmaceutical composition thereof.

In yet another method of treatment aspect, this invention providesmethods of treating a mammal susceptible to or afflicted with diseasesand disorders which are mediated by or result in inflammation such as,for example rheumatoid arthritis and osteoarthritis, myocardialinfarction, various autoimmune diseases and disorders, uveitis andatherosclerosis; itch/pruritus such as, for example psoriasis; and renaldisorders. Such method comprises administering an effectivecondition-treating or condition-preventing amount of one or more of thecompounds of the invention or pharmaceutical compositions justdescribed.

This invention also relates to the use of the present compounds in themanufacture of a medicament for treatment or prophylaxis of a conditionprevented, ameliorated or eliminated by administration of an inhibitorof Mitogen-Activated Protein Kinase-Activated Protein Kinase 5, or acondition characterised by abnormal collagenase activity, or a conditionassociated with ECM degradation or a condition selected from diseasesinvolving inflammation, most preferably in for the treatment ofrheumatoid arthritis.

As a further aspect of the invention there is provided the presentcompounds for use as a pharmaceutical especially in the treatment orprevention of the aforementioned conditions and diseases. Also providedherein is the use of the present compounds in the manufacture of amedicament for the treatment or prevention of one of the aforementionedconditions and diseases.

A preferred regimen of the present method comprises the administrationto a subject in suffering from a disease condition characterized asinflammatory, with an effective matrix metallo-protease inhibitingamount of a compound of the present invention for a period of timesufficient to reduce the abnormal levels of extracellular matrixdegradation in the patient, and preferably terminate, theself-perpetuating processes responsible for said degradation. A specialembodiment of the method comprises administering of an effective matrixmetallo-protease inhibiting amount of a compound of the presentinvention to a subject patient suffering from or susceptible to thedevelopment of rheumatoid arthritis, for a period of time sufficient toreduce or prevent, respectively, collagen and bone degradation in thejoints of said patient, and preferably terminate, the self-perpetuatingprocesses responsible for said degradation.

Injection dose levels range from about 0.1 mg/kg/hour to at least 10mg/kg/hour, all for from about 1 to about 120 hours and especially 24 to96 hours. A preloading bolus of from about 0.1 mg/kg to about 10 mg/kgor more may also be administered to achieve adequate steady statelevels. The maximum total dose is not expected to exceed about 2 g/dayfor a 40 to 80 kg human patient.

For the prevention and/or treatment of long-term conditions, such asinflammatory and autoimmune conditions, the regimen for treatmentusually extends over many months or years, and accordingly oral dosingis preferred for patient convenience and tolerance. With oral dosing,one to five and especially two to four and typically three oral dosesper day are representative regimens. Using these dosing patterns, eachdose provides from about 0.01 to about 20 mg/kg of the compound of theinvention, with preferred doses each providing from about 0.1 to about10 mg/kg and especially about 1 to about 5 mg/kg.

Transdermal doses are generally selected to provide similar or lowerblood levels than are achieved using injection doses.

When used to prevent the onset of an inflammatory condition, thecompounds of this invention will be administered to a patient at riskfor developing the condition, typically on the advice and under thesupervision of a physician, at the dosage levels described above.Patients at risk for developing a particular condition generally includethose that have a family history of the condition, or those who havebeen identified by genetic testing or screening to be particularlysusceptible to developing the condition.

The compounds of this invention can be administered as the sole activeagent or they can be administered in combination with other agents,including other compounds that demonstrate the same or a similartherapeutic activity, and that are determined to safe and efficaciousfor such combined administration.

GENERAL SYNTHETIC PROCEDURES

The imidazo[1,2-a]pyridyl compounds of this invention can be preparedfrom readily available starting materials using the following generalmethods and procedures. It will be appreciated that where typical orpreferred process conditions (i.e., reaction temperatures, times, moleratios of reactants, solvents, pressures, etc.) are given, other processconditions can also be used unless otherwise stated. Optimum reactionconditions may vary with the particular reactants or solvent used, butsuch conditions can be determined by one skilled in the art by routineoptimization procedures.

Additionally, as will be apparent to those skilled in the art,conventional protecting groups may be necessary to prevent certainfunctional groups from undergoing undesired reactions. The choice of asuitable protecting group for a particular functional group as well assuitable conditions for protection and deprotection are well known inthe art. For example, numerous protecting groups, and their introductionand removal, are described in T. W. Greene and P. G. M. Wuts, ProtectingGroups in Organic Synthesis, Second Edition, Wiley, New York, 1991, andreferences cited therein.

The following methods are presented with details as to the preparationof representative bicycloheteroaryls that have been listed hereinabove.The compounds of the invention may be prepared from known orcommercially available starting materials and reagents by one skilled inthe art of organic synthesis.

Synthetic Preparation of Compounds of the Invention

Compounds according to the present invention are produced according tothe following scheme.

General Scheme for Synthesising Compounds of Formula (I)

The C8 bromine of 5,8-dibromoimidazo[1,2-a]pyrazine (D) can beselectively displaced with primary and secondary amines or anilines asdescribed in the general reaction scheme. When (D) is displaced byanilines or primary amines (R¹═H, R⁸ for example, 4-chlorophenyl) theproduct compounds of the general formula (E) are protected as thet-butyl carbamate to give compounds of the general formula (F) which arereacted with the corresponding boronic acids (for example R3=4-pyrazole)to give the desired targets (I) after removal of the protecting group.

General Procedures:

Synthesis of compound (B) as described in the general reaction scheme,3,6-dibromo-pyrazine-2-carboxylic acid.

LiOH (655 mg, 27 mmol) is added to a solution ofmethyl-3-amino-2-pyrazine carboxylate (A) (J. Med. Chem. 1969, 12,285-87) (2.7 g, 9 mmol) in THF:water:MeOH (18:4.5:4.5 mL). The reactionis stirred at 5° C. for 30 min, concentrated in vacuo, taken up in DCMand washed with 1N HCl. The organic phase is dried over MgSO₄ andconcentrated in vacuo to afford compound (B). ¹H NMR (250 MHz,CDCl₃)δ(ppm)8.70(1H, s).

Synthesis of compound (C) as described in the general reaction scheme,3,6-Dibromo-pyrazin-2-ylamine.

Diphenylphosphorylazide (2.59 mL, 12 mmol) and triethylamine (1.67 mL,12 mmol) are added to a solution of 2,5-dibromo-3-pyrazoic acid (3.52 g,12 mmol) in t-butanol (90 mL). The reaction is heated at reflux for 18hours. The reaction is quenched with water, then concentrated in vacuoand taken up in DCM. The organic solution is washed with water and 1NNaOH, dried over MgSO₄ and concentrated in vacuo. The resultant solid isfiltered through a pad of silica using EtOAc, then concentrated andTFA:DCM (4:1, 12 mL) is added to the solid and stirred for 30 min. Thesolution is concentrated in vacuo then neutralised with 1N NaOH andextracted with DCM. The organic layer is dried over MgSO₄ andconcentrated in vacuo to give the product. ¹H NMR (250 MHz, d₆-DMSO)δ(ppm) 7.25 (2H, br s), 7.68 (1H, s); m/z (APCI) 254 (M+H)⁺; m.p135-139° C.

Synthesis of compound D as described in the general reaction scheme;5,8-Dibromoimidazo[1,2-a]pyrazine.

Bromoacetaldehyde diethyl acetal (49 mL, 326 mmol) and 48% hydrobromicacid is heated to reflux for 1.5 h, then poured into propan-2-ol (600mL) and quenched with NaHCO₃. After filtering,2,5-dibromo-3-aminopyrazine (41.34 g, 163 mmol) is added to the solutionand heated at reflux overnight. The reaction is cooled and solventsremoved in vacuo, followed by addition of aq. NaHCO₃ and extraction withEtOAc. The organic phase is dried over MgSO₄, filtered, and concentratedin vacuo to afford a brown solid. ¹H NMR (250 MHz, CDCl₃) δ(ppm)7.86(1H, s), 7.93-7.94 (1H, d), 7.98-7.99 (1H,d); m/z (APCI) 278 (M+H)⁺; m.p132-135° C.

Typical Example of Compound of Formula (E).

General procedure for amine displacement

Amine (1.5 eq., 10.8 mmol) and N,N-diisopropylethylamine (1.5 eq, 10.8mmol) are added to a solution of 5,8-dibromo-imidazo[1,2-a]pyrazine(1.99 g, 7.2 mmol) in ethanol (6 mL), and the reaction stirred at 80° C.for 15 hours. Ethanol is removed in vacuo and the product taken up inDCM and washed with water, dried over MgSO₄ and concentrated in vacuo.The resultant oil is passed through a pad of silica using DCM andconcentrated in vacuo to yield the desired product.

Typical Example of Compound of Formula (F)

General procedure for Boc protection

Di-tert-butyl-dicarbonate (3 eq, 10.8 mmol) andN,N-dimethylaminopyridine (0.1 eq, 0.36 mmol) are added to a solution ofthe (5-bromo-imidazo[1,2-a]pyrazin-8-yl)-amine (3.6 mmol) in DCM (4 mL).The solution is then heated at 50° C. overnight, then volatiles removedin vacuo and the product taken up in DCM and washed with 10% citric acidsolution. The organic layer is separated, dried over MgSO₄ and volatilesremoved in vacuo. Recrystallisation from DCM/hexane yields the desiredproduct.

Typical Example of Compound of Formula (I)

General procedure for Suzuki cross coupling reactions and Bocdeprotection

A solution of boronic acid in DMF (0.36 mmol, 0.6 mL) and 1.5MNa₂CO₃(aq.) solution (0.75 mmol, 0.5 mL) are added to a solution of (E)in DMF (0.3 mmol, 0.5 mL). Solutions of palladium acetate (95 mg),triphenylphosphine (335 mg) and catalyst (0.3 mL) in 1,4-dioxane areadded and the mixture is then heated at 80° C. for 16 hours. Forcompounds that require deprotection, the plates are evaporated todryness and resuspended in a 4:1 mixture of TFA:DCM (1 mL). The platesare agitated overnight and evaporated to dryness. The compounds areresuspended in DMF (2 mL) and purified by preparative reverse phaseHPLC.

Synthesis of Intermediates Intermediate 1:(5-Bromo-imidazo[1,2-a]pyrazin-8-yl)-(4-chloro-phenyl)-amine

Following the general procedure for amine displacement5,8-dibromo-imidazo[1,2-a]pyrazine (1.99 g, 7.2 mmol) and4-chloroaniline (1.37 g, 10.8 mmol) are coupled to give the titlecompound. Purification on silica gel with dichloromethane, methanol(98:2) gives the final product.

HPLC (254 nm): Rt 3.04 min (100%); m/z (APCI) 323, 325, 327 (M+H)⁺; ¹HNMR (250 MHz, CDCl₃) δ(ppm) 7.32-7.36 (2H, m), 7.56 (1H, s), 7.64 (1H,m), 7.76-7.80 (3H, m), 7.96 (1H, br s).

Intermediate 2:(5-Bromo-imidazo[1,2-a]pyrazin-8-yl)-(4-methoxy-benzyl)carbamic acidtert-butyl ester

Following the general procedure for Boc protection,(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-(4-methoxybenzyl)-amine (1.18 g,3.6 mmol) is treated with di-tert-butyldicarbonate (2.34 g, 10.68 mmol)to give the title compound; HPLC (254 nm): R_(t) 2.84 min (100%); m/z(APCI) 433 (M+H)⁺, 333 (M+H-BOC); ¹H NMR (250 MHz, CDCl₃) δ(ppm)1.38(9H, s), 3.74 (3H, s), 6.77 (2H, d), 7.33 (2H, d), 7.80-7.82 (3H, m).

Specific Synthetic Examples of Compounds of the Invention Compound 1:(4-Morpholin-4-yl-phenyl)-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine

Following the general procedure for Suzuki cross coupling and Bocdeprotection(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-(4-morpholin-4-yl-phenyl) carbamicacid tert-butyl ester (142 mg, 0.3 mmol) is coupled withpyrazole-4-boronic acid pinacol ester (70 mg, 0.36 mmol) to give thetitle compound. The reaction mixture is purified on silica gel usingdichloromethane/ methanol (90/10) eluant gave the title compound. HPLC(254 nm): Rt 2.16 min (100%); m/z (APCI) 362 (M+H)⁺; ¹H NMR (400 MHz,d₆-DMSO) δ(ppm)3.25 (4H, t), 3.94 (4H, t), 7.12 (2H, d), 7.73 (1H, s),7.88 (1H, d), 8.08 (2H, d), 8.20 (1H, s), 8.27 (1H, d), 8.54 (1H, s),9.49 (1H, s), 13.55 (1H, br s).

Compound 32:N-(2-Diethylamino-ethyl)-4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide

Step 1:4-(5-Bromo-imidazo[1,2-a]pyrazin-8-ylamino)-N-(2-diethylamino-ethyl)-benzamide

In the same way as described in for Compound 90, step 3, using4-(5-bromo-imidazo[1,2-a]pyrazin-8-ylamino)-benzoic acid (0.593 g 1.78mmol), N,N diethylethane-1,2-diamine (0.30 mL, 2.14 mmol), DIPEA (0.48mL, 2.68 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (0.68 g, 1.78 mmol) in DMF. The title compound (0.53g, 69%) is obtained after purification by silica gel columnchromatography eluting with DCM followed by 95:5 DCM:NH₃ (7M in MeOH).LCMS: Rt 2.21 min (96%).

Step 2:N-(2-Diethylamino-ethyl)-4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide

In the same way as described for Compound 85, step 1, using4-(5-bromo-imidazo[1,2-a]pyrazin-8-ylamino)-N-(2-diethylamino-ethyl)-benzamide(0.12 g, 0.28 mmol),4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole (65 mg,0.33 mmol), Pd(PPh₃)₄ (32 mg, 0.028 mmol) and NaO^(t)Bu (0.11 g, 1.11mmol) in DMF/water. Purification by silica gel column chromatographyeluting with 92:8 DCM:NH₃ (7M in MeOH) affords the title compound(0.0815 g, 70%). LCMS Rt 1.93 min (99%), m/z (APCI) 419 (M+H)⁺; ¹H-NMR(400 MHz, d₆-DMSO) δ(ppm) 0.82-0.95 (6H, t), 1.64 (2H, s) 2.37-2.43 (2H,m), 3.17-3.22 (4H, m), 6.55 (1H, br s), 7.20 (1H, br s), 7.55 (1H, s),7.65-7.69 (3H, m), 7.98-8.09 (4H, m), 9.67 (1H, s), 13.30 (1H, br s).

Compound 69:N-(2-Diethylaminoethyl)-4-[5-(1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-ylamino]benzenesulfonamide

Step 1: N-(2-Diethylaminoethyl)-4-nitrobenzenesulfonamide

A solution of 2-(N,N-diethyl)ethylamine (0.56 mL, 4.0 mmol) and pyridine(0.7 mL, 8.8 mmol) in DCM is cooled to 0° C. 4-Nitrobenzenesulfonylchloride (0.98 g, 4.4 mmol) is added and the solution stirred, whilstwarming to room temperature, for 18 hours. After this time the solutionis cooled to 0° C., and the resultant precipitate filtered, washed withDCM and air-dried. The title sulphonamide is isolated as a cream solid.

Step 2: 4-Amino-N-(2-diethylaminoethyl)benzenesulfonamide

A mixture of N-(2-diethylaminoethyl)-4-nitrobenzenesulfonamide (0.72 g,2.13 mmol) in cyclohexene (4.8 mL) and EtOH (24 mL) is heated using anoil bath until dissolution had almost been achieved. At this point Pd onC (144 mg, 10% (w/w)) is added, and the mixture heated at reflux for 5hours. After this time the solution is cooled to room temperature, thenfiltered through Hy-flo. The filtrate is evaporated in vacuo to afford ayellow gum. The yellow gum is dissolved in the minimum quantity ofmethanol, then NaHCO₃ (1 eq.) dissolved in water, is added. The mixtureis stirred at room temperature, then the mixture extracted with EtOAc(2×). The combined organic layers are dried (MgSO₄), and evaporated toafford the title aniline.

Step 3:4-(5-Bromoimidazo[1,2-a]pyrazin-8-ylamino)-N-(2-diethylaminoethyl)benzenestulfonamide

A mixture of 5,8-dibromoimidazo[1,2-a]pyrazine (1.0 g, 3.6 mmol),4-amino-N-(2-diethylaminoethyl)benzenesulfonamide (109 mg, 0.4 mmol),Pd₂(dba)₃ (7 mg, 0.007 mmol), Xantphos (8.4 mg, 0.015 mmol) and Cs₂CO₃(167 mg, 0.52 mmol) in dioxane is heated at 85° C. under nitrogen for 18hours. The reaction is cooled to room temperature then evaporated todryness. The residue is chromatographed on silica gel, eluting with DCMthen 98:2 DCM:NH₃ (7N in MeOH), and the fractions containing the titlecompound are combined and evaporated to afford a dark oil. This oilcontains an impurity, but is used in the subsequent step without furtherpurification.

Step 4:N-(2-Diethylaminoethyl)-4-[5-(1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-ylamino]benzenesulfonamide

In the same way as described for Compound 85, step 1, using4-(5-bromoimidazo[1,2-a]pyrazin-8-ylamino)-N-(2-diethylaminoethyl)benzenesulfonamide(100 mg, 0.21 mmol),4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole (17 mg,0.086 mmol), Pd(PPh₃)₄ (9 mg, 0.007 mmol) and NaO^(t)Bu (27 mg, 0.29mmol) in DMF:water (3:1, 6 mL). The crude residue is chromatographed onsilica gel, eluting with DCM followed by 97:3 DCM:NH₃ (7M in MeOH). Thefractions containing the desired product are combined and evaporated, toafford the title compound as a yellow solid. HPLC (254 nm): Rt 2.04 min(97.2%); m/z (APCI) 455 (M+H)⁺

Compound 85:4-[8-(4-Morpholin-4-yl-phenylamino)imidazo[1,2-a]pyrazin-5-yl]-1H-pyridin-2-one

Step 1:[5-(2-Ethoxypyridin-4-yl)imidazo[1,2-a]pyrazin-8-yl]-(4-morpholino-4-ylphenyl)amine

A mixture of2-ethoxy-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)pyridine (2.84g, 11.4 mmol),(5-bromoimidazo[1,2-a]pyrazin-8-yl)-4-(4-morpholino-4-ylphenyl)amine(0.85 g, 2.28 mmol), Pd(PPh₃)₄ (263 mg, 0.022 mmol) and NaO^(t)Bu (0.88g, 9.1 mmol) in DMF (20 mL) and water (7mL) is degassed using nitrogen,then heated at 85° C. for 18 hours. After this time the reaction mixtureis cooled to room temperature and the solvents removed in vacuo. Theresidue is chromatographed on silica gel, eluting with DCM followed by98:2 DCM:NH₃ (7M in MeOH), and the fractions containing the desiredproduct are combined and evaporated to afford the title ethoxypyridineas a solid. This material is contaminated with some de-brominatedcompound, and is used without further purification in the next step.

Step 2:(4-Morpholin-4-yl)phenyl)-[5-(2-hydroxypyridin-4-yl)imidazo[1,2-a]pyrazin-8-yl]amine

A mixture of[5-(2-ethoxypyridin-4-yl)imidazo[1,2-a]pyrazin-8-yl]-(4-morpholino-4-ylphenyl)amine(1.44 g, 3.46 mmol) and pyridinium hydrochloride (2 g, 17.3 mmol) inwater (0.5 mL) is heated at 150° C. for 1 hour in a sealed tube. Afterthis time the solvent is removed in vacuo. The residue ischromatographed on silica gel, eluting with DCM followed by 95:5 DCM:NH₃(7M in MeOH), and the fractions containing the desired product arecombined and evaporated. The residue is triturated with Et₂O to affordthe title compound as a solid. HPLC (254 nm): Rt 2.21 min (99.1%); m/z(APCI) 389 (M+H)⁺.

Compound 90N-(4-Hydroxybenzyl)-4-[5-(1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-ylamino]benzamide

Step 1: 4-(5-Bromoimidazo[1,2-a]pyrazin-8-ylamino) benzoic acid methylester

A mixture of 5,8-dibromoimidazo[1,2-a]pyrazine (2 g, 7.3 mmol), 4-aminobenzoic acid methyl ester (0.93 g, 6.2 mmol), NaO^(t)Bu (0.98 g, 10.2mmol), Pd₂(dba)₃ (133 mg, 0.14 mmol), Xantphos (168 mg, 0.29 mmol) andtoluene is degassed with nitrogen, then heated at 85° C. for 18 hours.The toluene is removed in vacuo, then MeOH is added to the cruderesidue. The solid is collected by filtration and dried in the vacuumoven. The solid is identified as the desired title ester. The filtrateis chromatographed with petrol:EtOAc (70:30 followed by 50:50) and thefractions containing the desired product, combined, evaporated andtriturated with MeOH. The resultant solid is collected by filtration andis also identified as the desired ester.

Step 2: 4-(5-Bromoimidazo[1,2-a]pyrazin-8-ylamino) benzoic acid

A solution of 4-(5-bromoimidazo[1,2-a]pyrazin-8-ylamino) benzoic acidmethyl ester (6.02 g, 17.3 mmol) and LiOH (7.3 g, 174 mmol) in THF (200mL) and water (200 mL) is stirred at 25° C. for 18 hours. The majorityof the solvents are removed in vacuo, then to the remaining aqueoussolution is added EtOAc. The aqueous layer is separated, then washedonce more EtOAc. The aqueous phase is separated, acidified to pH 5 usingconcentrated hydrochloric acid and the resultant solid filtered anddried in the vacuum oven. The solid is identified as the desired titleacid.

Step 3: 4-(5-Bromoimidazo[1,2-a]pyrazin-8-ylamino)-N-(4-hydroxybenzyl)benzamide

A mixture of 4-(5-bromoimidazo[1,2-a]pyrazin-8-ylamino) benzoic acid(116 mg, 0.35 mmol), 4-aminomethylphenol (43 mg, 0.35 mmol), andN,N-diisopropylethylamine (0.094 mL, 0.525 mmol) in DMF (1 mL) is cooledto 0° C., and O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (160 mg, 0.42 mmol) is added. The cooling bath isremoved, and the reaction mixture stirred at 25° C. for 18 hours. Afterthis time the solvent is removed in vacuo, and the residue trituratedwith MeOH. The resultant solid is collected by filtration and identifiedas the desired phenol.

Step 4:N-(4-Hydroxybenzyl)-4-[5-(1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-ylamino]benzamide

In the same way as described for Compound 85, step 1, using4-(5-bromoimidazo[1,2-a]pyrazin-8-ylamino)-N-(4-hydroxybenzyl) benzamide(47 mg, 0.11 mmol),4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole (25 mg,0.13 mmol), Pd(PPh₃)₄ (12 mg, 0.011 mmol) and NaO^(t)Bu (41 mg, 0.43mmol) in DMF/water (3:1). The crude residue is chromatographed on silicagel, eluting with petrol:EtOAc (50:50 followed by 0:100) and thenEtOAc:MeOH (95:5). The fractions containing the desired product arecombined and evaporated, to afford the title compound as a solid. HPLC(254 nm): Rt 2.38 min (90.9%); m/z (APCI) 426 (M+H)⁺.

Compound 97:4-[8-(4-Methanesulfonyl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-benzamide

Step 1:(5-Bromo-imidazo[1,2-a]pyrazin-8-yl)-(4-methanesulfonyl-phenyl)-amine

A degassed mixture of 5,8-dibromo-imidazo[1,2-a]pyrazine (2.19 g, 7.916mmol), 4-methylsulfonylaniline (1.49 g, 8.708 mmol), Pd₂dba₃ (145 mg,0.15 mmol) and Xantphos (183 mg, 0.317 mmol), in dry toluene (50 mL) isstirred at 110° C. for 16 hours. After evaporation of the solvent, theresidue is purified by silica gel column chromatography eluting with95:5 DCM:NH₃ (7M in MeOH). The title compound (1.496 g, 51%) is isolatedcontaining some starting material (20%) and used in the next stepwithout further purification.

Step 2:4-[8-(4-Methanesulfonyl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-benzamide

In the same way as described for Compound 85, step 1, using4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzamide (1.0 g, 6.11mmol),(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-(4-methanesulfonyl-phenyl)-amine(1.49 g, 4.07 mmol), Pd(PPh₃)₄ (471 mg, 0.41 mmol) and NaO^(t)Bu (1.57g, 0.82 mmol) in DMF (18 mL) and water (6 mL). The residue ischromatographed on silica gel, eluting with DCM followed by 95:5 DCM:NH₃(7M in MeOH), and the fractions containing the desired product arecombined and evaporated to afford a solid, crystallised in ethyl acetate(566 mg, 34%). LCMS: Rt 2.53 min (99%), m/z (APCI) 408 (M+H)⁺; ¹H-NMR(400 MHz, d₆-DMSO δ(ppm) 3.21 (3H, s), 7.54 (1H, br s), 7.67 (1H, s),7.81 (1H, s), 7.86-7.92 (4H, m), 8.09-8.17 (3H, m), 8.21 (1H, br s),8.41 (2H, d), 10.32 (1H, s).

Compound 102:4-{8-[4-(4-Methylpiperazin-1-yl)-phenylamino]imidazo[1,2-a]pyrazin-5-yl}-1H-pyridin-2-one

Step 1:(4-(4-Methylpiperazin-1-yl)phenyl)-[5-(2-ethoxypyridin-4-yl)imidazo[1,2-a]pyrazin-8-yl]amine

A mixture of2-ethoxy-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)pyridine (97mg, 0.31 mmol),(5-bromoimidazo[1,2-a]pyrazin-8-yl)-(4-(4-methylpiperazin-1-yl)phenyl)amine(100 mg, 0.259 mmol), Pd(PPh₃)₄ (29 mg, 0.026 mmol) and NaO^(t)Bu (99mg, 1.0 mmol) in DMF (3 mL) and water (1 mL) is heated at 85° C. for 18hours. After this time the reaction mixture is cooled to roomtemperature and the solvents removed in vacuo. The residue ischromatographed on silica gel, eluting with DCM followed by 96:4 DCM:NH₃(7M in MeOH), and the fractions containing the desired product arecombined and evaporated to afford the title ethoxypyridine as a solid.

Step 2:4-{8-[4-(4-Methylpiperazin-1-yl)phenylamino]imidazo[1,2-a]pyrazin-5-yl}-1H-pyridin-2-one

A mixture of(4-(4-methylpiperazin-1-yl)phenyl)-[5-(2-ethoxypyridin-4-yl)-imidazo[1,2-a]pyrazin-8-yl]amine(103 mg, 0.24 mmol) and pyridinium hydrochloride (130 mg, 1.2 mmol) inwater (0.2 mL) is heated at 150° C. for 25 minutes in a sealed tube.After this time the solvent is removed in vacuo. The residue ischromatographed on silica gel, eluting with DCM followed by 92:8 DCM:NH₃(7M in MeOH), and the fractions containing the desired product arecombined and evaporated to afford the title compound as a solid. HPLC(254 nm): Rt 1.70 min (99.7%); m/z (APCI) 402 (M+H)⁺.

Compound 103:5-[5-(1H-Pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-pyridine-2-carboxylicacid (2-diethylamino-ethyl)-amide

Step 1: 5-Amino-pyridine-2-carboxylic acid (2-diethylamino-ethyl)-amide

5-Aminopyridine-2-carbonitrile (0.2 g, mmol) is stirred in a mixture ofEtOH (1 mL) and 48% aq NaOH (1 mL) at 120°C. overnight. The solvent isremoved in vacuo and the water residue is washed with EtOAc. The waterlayer is acidified to pH 5 and concentrated to afford the crude5-amino-pyridine-2-carboxylic acid (0.23 g, 0.168 mmol).

The 5-amino-pyridine-2-carboxylic acid is coupled toN,N-diethyl-ethane-1,2-diamine (0.47 mL, 3.32 mmol), in the presence ofO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (1.26 g, 3.33 mmol), and DIPEA (0.59 mL, 3.33 mmol)in THF (10 mL). The mixture is stirred at room temperature for 48 hours.The solvent is removed in vacuo and the residue partitioned betweenwater-EtOAc. The organic layer is dried over MgSO₄, filtered andconcentrated to give a residue purified by silica gel columnchromatography. 5-Amino-pyridine-2-carboxylic acid(2-diethylamino-ethyl)-amide is isolated (394 mg, 100%) eluting with96:4 DCM:NH₃ (7M in MeOH).

Step 2:5-(5-Bromo-imidazo[1,2-a]pyrazin-8-ylamino)-pyridine-2-carboxylic acid(2-diethylamino-ethyl)-amide

In the same way as described for Compound 90, step 1, using5,8-dibromoimidazo[1,2-a]pyrazine (0.272 g, 0.982 mmol),5-amino-pyridine-2-carboxylic acid (2-diethylamino-ethyl)-amide (0.197g, 0.835 mmol), NaO^(t)Bu (0.132 g, 1.37 mmol), Pd₂(dba)₃ (18 mg, 0.02mmol), Xantphos (22 mg, 0.038 mmol) and toluene (3 mL). The crudematerial is purified by silica gel column chromatography eluting withDCM followed by 94:6 DCM:NH₃ (7M in MeOH) to afford the title compound(0.263 g, 72%). HPLC (254 nm) Rt 2.09 min (72%).

Step 3:5-[5-(1H-Pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-pyridine-2-carboxylicacid (2-diethylamino-ethyl)-amide

In the same way as described for Compound 85, step 1, using5-(5-bromo-imidazo[1,2-a]pyrazin-8-ylamino)-pyridine-2-carboxylic acid(2-diethylamino-ethyl)-amide (169 mg, 0.39 mmol),4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole (91 mg,0.47 mmol), Pd(PPh₃)₄ (45 mg, 0.039 mmol) and NaO^(t)Bu (150 mg, 1.56mmol) in DMF:water (3:1, 9 mL). The crude residue is purified by reversephase preparative HPLC to give the title compound (30 mg, 18%). LCMS: Rt1.81 min (97.6%); m/z (APCI) 420 (M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO) δ(ppm). 1.02 (6H, t), 2.36-2.71 (4H, m), 3.09-3.40 (4H, m), 7.73 (1H, s),7.84 (1H, s), 8.04 (1H, d), 8.24 (3H, m), 8.56 (1H, m), 8.79 (1H, d),9.28 (1H, s), 10.24 (1H, s), 13.40 (1H, br s).

Compound 108:4-[5-(2-Oxo-1,2-dihydropyridin-4-yl)imidazo[1,2-a]pyrazin-8-ylamino]-N-pyridin-3-ylmethylbenzamide

Step 1: 4-(5-Bromoimidazo[1,2-a]pyrazin-8-ylamino)-N-pyridin-3-ylmethylbenzamide

In the same way as described for Compound 90, step 3, using4-(5-bromoimidazo[1,2-a]pyrazin-8-ylamino) benzoic acid (365 mg, 1.57mmol), 3-aminomethylpyridine (0.16 mL, 1.57 mmol),N,N-diisopropylethylamine (0.42 mL, 2.35 mmol),O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (HATU) (715 mg, 1.9 mmol) and DMF (6mL). The cruderesidue is purified using column chromatography, eluting with EtOAcfollowed by EtOAc:MeOH (95:5) then EtOAc:NH₃ (7M in MeOH). The titleamide is isolated as a solid.

Step 2:4-[5-(2-Ethoxypyridin-4-yl)imidazo[1,2-a]pyrazin-8-ylamino]-N-pyridin-3-ylmethylbenzamide

A mixture of2-ethoxy-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)pyridine (85mg, 0.34 mmol),4-(5-bromoimidazo[1,2-a]pyrazin-8-ylamino)-N-pyridin-3-ylmethylbenzamide (150 mg, 0.35 mmol), Pd(PPh₃)₄ (33 mg, 0.028 mmol) andNaO^(t)Bu (109 mg, 1.1 mmol) in DMF:water (3:1, 9 mL) is heated at 85°C. for 18 hours. After this time the reaction mixture is cooled to roomtemperature and the solvents removed in vacuo. The residue ischromatographed on silica gel, eluting with DCM followed by 95:5 DCM:NH₃(7M in MeOH), and the fractions containing the desired product arecombined and evaporated to afford the title ethoxypyridine as a solid.

Step 3:4-[5-(2-Oxo-1,2-dihydropyridin-4-yl)imidazo[1,2-a]pyrazin-8-ylamino]-N-pyridin-3ylmethylbenzamide

A mixture of4-[5-(2-ethoxypyridin-4-yl)imidazo[1,2-a]pyrazin-8-ylamino]-N-pyridin-3-ylmethylbenzamide (82 mg, 0.18 mmol) and pyridinium hydrochloride (101 mg, 0.88mmol) in water (0.5 mL) is heated at 150° C. for 20 minutes in a sealedtube. After this time the solvent is removed in vacuo. The residue ischromatographed on silica gel, eluting with DCM followed by 90:10DCM:NH₃ (7M in MeOH), and the fractions containing the desired productare combined and evaporated. The title compound is isolated as a solid.HPLC (254 nm): Rt 1.83 min (99.1%); m/z (APCI) 438 (M+H)⁺.

Compound 111:[5-(2-Dimethylamino-pyridin-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-(4-morpholin-4-yl-phenyl)-amine

In the same way as described for Compound 85, step 1, using2-fluoropyridine-4-boronic acid (0.047 g, 0.337 mmol),5-bromo-imidazo[1,2-a]pyrazin-8-yl)-4-morpholin-4-yl-phenylamine (0.105g, 0.281 mmol), Pd(PPh₃)₄ (32 mg, 0.0281 mmol) and NaO^(t)Bu (0.108 g,1.12 mmol) in N,N-dimethylformamide/water (7 mL). The residue ischromatographed on silica gel, eluting with DCM followed by 95:5 DCM:NH₃(7M in MeOH), and the fractions containing the desired product arecombined and evaporated to afford the title compound containing someside products. Additional purification by reverse phase prep HPLCaffords the title compound as a yellow solid (18 mg, 16%). LCMS: Rt 1.99min (98%), m/z (APCI) 416 (M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO) δ (ppm)3.09-3.13 (10H, m), 3.78 (4H, m), 6.89 (2H, m), 6.97 (2H, d), 7.59 (1H,s), 7.72 (1H, s), 7.93 (2H, d), 8.09 (1H, s), 8.12 (1H, d), 9.60 (1H,s).

Compound 119:(4-(4-Methylpiperazin-1-yl)phenyl)-[5-(5-methyl-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-yl]amine

In the same way as described for Compound 127, step 4, using5-methyl-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole(75 mg, 0.36 mmol),(5-bromoimidazo[1,2-a]pyrazin-8-yl)-(4-(4-methylpiperazin-1-yl)phenyl)amine(70 mg, 0.18 mmol) and Pd(PPh₃)₄ (52 mg, 0.045 mmol) in dioxane (0.6mL), DMF (1.33 mL) and 1.5M Na₂CO₃ (0.97 mL). The residue ischromatographed on silica gel, eluting with DCM followed by 95:5 DCM:NH₃(7M in MeOH), and the fractions containing the desired product arecombined and evaporated. The title compound is isolated as a solid. HPLC(254 nm): Rt 1.62 min (98.2%); m/z (APCI) 389 (M+H)⁺.

Compound 121:5-{8-[4-(4-Methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-nicotinamide

Step 1: 5-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-nicotinamide

In the same way as described for Compound 209, step 2, using5-bromo-nicotinamide (0.2 g, 0.99 mmol), bis(pinacolato)diboron (0.27 g,1.09 mmol), PdCl₂dppf (0.024 g, 0.029 mol) and KOAc (0.29 g, 2.98 mmol)suspended in dioxane (2 mL). The crude compound is used in the next stepwithout further purification.

Step 2:5-{8-[4-(4-Methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-nicotinamide

In the same way as described for Compound 127, step 4, using5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-nicotinamide (51.6 mg,0.207 mmol),(5-bromoimidazo[1,2-a]pyrazin-8-yl)-(4-(4-methylpiperazin-1-yl)phenyl)amine(40 mg, 0.103 mol), Pd(PPh₃)₄ (30 mg, 0.026 mmol) and 1.5M Na₂CO₃ (0.55mL), in dioxane (0.46 mL) and DMF (1.01 mL). The residue ischromatographed on silica gel, eluting with DCM followed by 96:4 DCM:NH₃(7M in MeOH) to afford the title compound (4.8 mg, 11%). LCMS: Rt 1.74min (97%) m/z (APCI) 429; ¹H-NMR (400 MHz, d₆-DMSO) δ (ppm).2.26 (3H,s), 2.49-2.54 (4H, m), 3.12 (4H, m), 6.97 (2H, d), 7.57 (1H, s), 7.73(1H, s), 7.75 (1H, br s), 7.91 (2H, d), 8.07 (1H, s), 8.29 (1H, br s),8.54 (1H, s), 9.03 (1H, s), 9.14 (1H, s), 9.58 (1H, s).

Compound 122:3-Fluoro-4-{8-[4-(4-methylpiperazin-1-yl)phenylamino]imidazo[1,2-a]pyrazin-5-yl}benzamide

In the same way as described for Compound 127, step 4, using3-fluoro-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl) benzamide (82mg, 0.31 mmol),(5-bromoimidazo[1,2-a]pyrazin-8-yl)-(4-(4-methylpiperazin-1-yl)phenyl)amine(60 mg, 0.155 mmol) and Pd(PPh₃)₄ (45 mg, 0.038 mmol) in dioxane (0.68mL), DMF (1.52 mL) and 1.5M Na₂CO₃ (0.83 mL). The residue ischromatographed on silica gel, eluting with DCM followed by 95:5 DCM:NH₃(7M in MeOH), and the fractions containing the desired product arecombined and evaporated. The title compound is isolated as a solid. HPLC(254 nm): Rt 1.92 min (95.0%); m/z (APCI) 446 (M+H)⁺.

Compound 123:2-Fluoro-4-{8-[4-(4-methylpiperazin-1-yl)phenylamino]imidazo[1,2-a]pyrazin-5-yl}benzamide

In the same way as described for Compound 127, step 4, using2-fluoro-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl) benzamide (82mg, 0.31 mmol),(5-bromoimidazo[1,2-a]pyrazin-1-yl)-(4-(4-methylpiperazin-1-yl)phenyl)amine(60 mg, 0.155 mmol) and Pd(PPh₃)₄ (45 mg, 0.038 mol) in dioxane (2.2 mL)and 1.5M Na₂CO₃ (0.83 mL). The residue is chromatographed on silica gel,eluting with DCM followed by 96:4 DCM:NH₃ (7M in MeOH), and thefractions containing the desired product are combined and evaporated.The residue is triturated with Et₂O to afford the title compound as asolid. HPLC (254 nm): Rt 1.94 min (93.5%); m/z (APCI) 446 (M+H)⁺

Compound 125:[5-(2-Fluoro-pyridin-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-[4-(4-methyl-piperazin-1-yl)-phenyl]-amine

In the same way as described for Compound 127, step 4, using2-fluoro-pyridine-4-boronic acid (44 mg, 0.310 mmol),(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-(4-(4-methylpiperazin-1-yl)phenyl)amine(60 mg, 0.155 mol), Pd(PPh₃)₄ (45 mg, 0.038 mmol) and 1.5M Na₂CO₃ (0.83mL) in dioxane (2.2 mL). The residue is chromatographed on silica gel,eluting with DCM followed by 97:3 DCM:NH₃ (7M in MeOH) affording thetitle compound (37.8 mg, 61%). LCMS: Rt 2.14 min (96%) m/z (APCI) 404(M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO) δ (ppm)2.26 (3H, s), 2.49-2.54 (4H, t)3.13-3.15 (4H, m), 6.97 (2H, d), 7.58 (1H, s), 7.71 (1H, s), 7.75 (2H,m), 7.89 (2H, d), 8.21 (1H, s), 8.43 (1H, d), 9.73 (1H, s).

Compound 126: 2-Dimethylamino-4-{8-[4-(4-methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide

Step 1:2-Fluoro-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzamide

In the same way as described for Compound 209, step 2, using4-bromo-2-fluoro-benzamide, (0.2 g, 0.94 mmol) bis(pinacolato)diboron(0.26 g, 1.01 mmol), PdCl₂dppf (0.023 g, 0.027 mol) and KOAc (0.29 g,2.76 mmol) in dioxane (2 mL). The crude compound is used in the nextstep without further purification.

Step 2:2-Dimethylamino-4-{8-[4-(4-methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide

In the same way as described for Compound 127, step 4, using2-fluoro-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzamide (148mg, 0.414 mmol),(5-bromoimidazo[1,2-a]pyrazin-8-yl)-(4-(4-methylpiperazin-1-yl)phenyl)amine(80 mg, 0.279 mol) and Pd(PPh₃)₄ (81 mg, 0.069 mmol) and 1.5M Na₂CO₃(1.49 mL), in dioxane (0.93 mL) and N,N-dimethylformamide (2.06 mL). Theresidue is chromatographed on silica gel, eluting with DCM followed by96:4 DCM:NH₃ (7M in MeOH). The compound, isolated after additionalpurification by reverse phase prep HPLC is2-dimethylamino-4-{8-[4-(4-methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide(3.2 mg). LCMS: Rt 1.67 min (97%) m/z (APCI) 471 (M+H)⁺; ¹H-NMR (400MHz, d₆-DMSO) δ (ppm).2.26 (3H, s), 2.48-2.53 (4H, m), 2.85 (6H, s),3.12-3.14 (4H, m), 6.96 (2H, d), 7.32 (1H, d), 7.36 (1H, d), 7.52 (1H,s), 7.54 (1H, br s), 7.71 (1H, s), 7.74 (1H, d), 7.91 (2H, d), 8.02 (1H,s), 8.28 (1H, br s), 9.48 (1H, s).

Compound 127:(4-(4-Methylpiperazin-1-yl)phenyl)-[5-(2-carboxamido-5-thienyl)imidazo[1,2-a]pyrazin-8-yl]amine

Step 1: 5-Bromo-thiophene-2-carboxylic acid amide

A solution of 5-bromo-thiophene-2-carboxylic acid (4.51 g, 21.78 mmol),3-hydroxybenzotriazole hydrate (3.24 g, 23.96 mmol),1-ethyl-3-(3′-dimethylaminopropyl)-carbodiimide (4.6 g, 23.96 mmol) inDMF (70 mL) is stirred at room temperature for 2 hours. The reactionmixture is then cooled to 0° C. and aq. 35% NH₃ (2.2 mL) is added. Themixture is stirred at room temperature overnight. The solvent is removedin vacuo and the residue dissolved in EtOAc, washed with 1N NaHCO3, andbrine. The organic layers are combined, dried over MgSO₄, filtered andconcentrated to afford the title compound (3.78 g, 84%). HPLC (254 nm):Rt 2.46 min (96.5%).

Step 2:5-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-thiophene-2-carboxylicacid amide

5-Bromo-thiophene-2-carboxylic acid amide (0.5 g, 2.426 mmol),bis(pinacolato)diboron (678 mg, 2.669 mmol), PdCl₂dppf (59 mg, 0.072mmol) and KOAc (0.714 g, 7.28 mmol) are suspended in dioxane (5 mL),purged with nitrogen for 5 minutes and then heated at 85° C. overnight.The solvent is removed in vacuo and the residue partitioned betweenethyl acetate and water. The aqueous layer is extracted again with ethylacetate and the combined organic phases are washed with brine, filteredthrough MgSO₄ and evaporated in vacuo to afford the title compound (417mg, 68%)

Step 3:(5-Bromoimidazo[1,2-a]pyrazin-8-yl)-(4-(4-methylpiperazin-1-yl)phenylamine

4-(4-Methylpiperazin-1yl)phenylamine (5.08 g, 26.6 mmol) andN,N-diisopropylethylamine (4.6 mL, 26.6 mmol) and5,8-dibromoimidazo[1,2-a]pyrazine (6.1 g, 22.2 mmol) in iso-propanol(150 mL), is heated at 90° C. for 2 days. The solvent is removed invacuo, and the crude residue partitioned between DCM and 1N NaOH. Theorganic phase is separated, then washed with water followed by brine.The organic layer is separated, dried (MgSO₄) and evaporated. The cruderesidue is chromatographed on silica gel, eluting with DCM followed by96:4 DCM:NH₃ (7M in MeOH), and the fractions containing the desiredproduct are combined and evaporated. The residue is triturated with Et₂Oto afford the title compound as a solid.

Step 4:(4-(4-Methylpiperazin-1-yl)phenyl)-[5-(2-carboxamido-5-thienyl)imidazo[1,2-a]pyrazin-8-yl]amine

A mixture of5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)thiophene-2-carboxylicacid amide (0.19 g, 0.75 mmol),(5-bromoimidazo[1,2-a]pyrazin-8-yl)-(4-(4-methylpiperazin-1-yl)phenyl)amine(60 mg, 0.155 mmol) and Pd(PPh₃)₄ (45 mg, 0.038 mmol) in dioxane (2.2mL) and 1.5M Na₂CO₃(0.83 mL) is heated at 85° C. for 18 hours. Afterthis time the reaction mixture is cooled to room temperature andfiltered. The filtrate is partitioned between EtOAc and water, and theorganic layer separated. The organic phase is washed with water (4×)then separated. The aqueous phase is then washed with EtOAc and theorganic layer separated. The combined organic layers are dried (MgSO₄)and evaporated. The residue is chromatographed on silica gel, elutingwith DCM followed by 95:5 DCM:NH₃ (7M in MeOH), and the fractionscontaining the desired product are combined and evaporated to afford thetitle compound as a solid. HPLC (254 nm): Rt 1.95 min (98.9%); m/z(APCI) 434 (M+H)⁺.

Compound 129:5-{8-[4-(4-Methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-pyridine-2-carboxylicacid amide

Step 1: 5-Tributylstannanyl-pyridine-2-carboxylic acid amide

A degassed mixture of 5-bromo-pyridine-2-carboxylic acid (0.2 g, 1mmol), tributyltin (1.16 g, 2 mmol) and PdCl₂(PPh₃)₂ (0.07 g, 0.1 mmol)in DMF (4 mL) is stirred at 115° C. Additional 20% of PdCl₂(PPh₃)₂ (0.14g, 0.2 mmol) is added and the reaction stirred for 24 hours. Thereaction mixture is partitioned between EtOAc and water, the organiclayer is washed with water (4×), dried over MgSO₄, filtered andconcentrated in vacuo. The crude is purified by silica gel columnchromatography eluting with 9:1 petroleum ether-ethyl acetate to affordthe title compound (0.132 mg, 32%).

Step 2:5-{8-[4-(4-Methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-pyridine-2-carboxylicacid amide

A degassed mixture of(5-bromoimidazo[1,2-a]pyrazin-8-yl)-(4-(4-methylpiperazin-1-yl)phenyl)amine(59 mg, 0.154 mol), 5-tributylstannanyl-pyridine-2-carboxylic acid amide(0.127, 0.309 mmol), PdCl₂(PPh₃)₂ (0.027 g, 0.038 mmol) in DMF (2 mL),is stirred at 85° C. for 20 hours. The reaction mixture is partitionedbetween EtOAc and water. The organic layer is washed with water (4×),dried over MgSO₄, filtered and concentrated in vacuo. The crude ispurified by silica gel column chromatography eluting with DCM followedby 97:3 DCM:NH₃ (7M in MeOH) affording the title compound (2.0 mg, 3%).LCMS: Rt 1.75 min (92%) m/z (APCI) 429 (M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO)δ (ppm) 2.26 (3H, s), 2.49-2.51 (4H, t), 3.12-3.15 (4H, m), 6.97 (2H,d), 7.62 (1H, s), 7.73 (1H, s), 7.77 (1H, br s), 7.91 (2H, d), 8.08 (1H,s), 8.22 (1H, d), 8.24 (1H, br s), 8.37-8.40 (1H, m), 8.96 (1H, s), 9.61(1H, s).

Compound 130:4-[3-Methyl-8-(4-morpholin-4-ylphenylamino)imidazo[1,2-a]pyrazin-5-yl]benzamide

This compound may be prepared using methods as described for Compound131, using 4-carbamoylbenzene boronic acid in Step 4. LCMS: Rt=1.44 min(100%), m/z (ESI) 429 (M+H)⁺.

Compound 131:[3-Methyl-5-(1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-yl]-(4-morpholin-4yl-phenyl)amine

Step 1: 2-Bromo-1,1-dimethoxypropane

A solution of propanal (12.5 mL, 170 mmol) in MeOH (85 mL) is stirred atrt and molecular sieves (3 Å, powdered, 4.3 g) are added. The mixture isheated at reflux, Br₂ (8.8 mL, 170 mmol) is added dropwise over 30 minand reflux is then continued for 4.5 h. The mixture is stirred overnightat rt and K₂CO₃ (11.9 g, 86 mmol) is then added and the resulting slurryis stirred for 2.5 h. This is filtered and the solid is washed with MeOH(20 mL). Brine (100 mL) is added to the filtrate and the mixture isextracted with pentane (3×75 mL). The extracts are dried over MgSO₄ andevaporated under reduced pressure to afford a pale yellow oil (16.0 g)which is not purified further.

Step 2: 5,8-Dibromo-3-methylimidazo[1,2-a]pyrazine

2,5-Dibromo-3-aminopyrazine (2.0 g, 7.9 mmol),2-bromo-1,1-dimethoxypropane (7.25 g, 40.0 mmol) and pyridiniump-toluenesulfonate (2.0 g, 7.9 mmol) are stirred in acetonitrile (65 mL)at reflux for 3 days. The mixture is cooled and the solvent evaporatedunder reduced pressure. The residue is partitioned between DCM (150 mL)and water (50 mL) and the layers separated. The organic fraction iswashed with NaHCO₃ (sat. aq., 50 mL) and brine (50 mL) and dried overMgSO₄. Evaporation of the solvent under reduced pressure gives a viscousblack oil (2.14 g) which is purified by silica chromatography, elutingwith 10% -20% EtOAc in cyclohexane to afford the title compound as ared-brown solid (280 mg, 1 mmol).

Step 3:(5-Bromo-3-methylimidazo[1,2-a]pyrazin-8-yl)-(4-morpholin-4-ylphenyl)amine

5,8-Dibromo-3-methylimidazo[1,2-a]pyrazine (270 mg, 0.93 mmol) and4-morpholinoaniline (200 mg, 1.1 mmol) are combined in nBuOH (10 mL) andDIPEA (240 μL, 1.4 mmol) is added. The mixture is heated at reflux for10 h, cooled and the solvent removed under reduced pressure. The residueis purified by silica chromatography, eluting with 12% -100% EtOAc incyclohexane, to afford the title compound as a red-brown solid (190 mg).

Step 4:[3-Methyl-5-(1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-yl]-(4-morpholin-4-yl-phenyl)amine

This compound may be prepared using methods as described for Compound184, step 4. LCMS: Rt=0.81 min (100%), m/z (ESI) 376 (M+H)⁺.

Compound 132:[5-(2-Amino-pyridin-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-[4-(4-methyl-piperazin-1-yl)-phenyl]-amine

A solution of[5-(2-fluoro-pyridin-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-[4-(4-methyl-piperazin-1-yl)-phenyl]-amine(0.0272, 0.067 mmol) (from Example 125) in a mixture of aq. NH₃ (2.5mL):1M NH₃ in MeOH (2.5 mL) is stirred at 100° C. overnight. Additionalaq. NE₃ (5 mL) and 0.5M NH₃ in dioxane (2 mL) are added and the mixturestirred at 120° C. for 4 days. The mixture is concentrated in vacuo andpurified by reverse phase HPLC affording the title compound (4.2 mg,16%). LCMS: Rt 3.76 min (95%) m/z (APCI) 401 (M+H)⁺; ¹H-NMR (400 MHz,d₆-DMSO) δ (ppm). 2.26 (3H, s), 2.49-2.53 (4H, t), 3.12-3.15 (4H, m),6.18 (2H, br s) 6.76 (1H, s), 6.81 (1H, d), 6.97 (2H, d), 7.51 (1H, s),7.72 (1H, s), 7.88 (2H, d), 8.04 (1H, s), 8.09 (1H, d), 9.55 (1H, s).

Compound 136:4-{8-[3-Chloro-4-(4-methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide

Step 1:(5-Bromo-imidazo[1,2-a]pyrazin-8-yl)-[3-chloro-4-(4-methyl-piperazin-1-yl)-phenyl]-amine

Following the general procedure for amine displacement using5,8-dibromoimidazo[1,2-a]pyrazine (0.20 g, 0.72 mmol),3-chloro-4-(4-methyl-piperazin-1-yl)-phenylamine (0.263 g, 1.08 mmol)and DIPEA (0.19 mL, 1.08 mmol) in ^(i)PrOH (5 mL). Purification of thecrude compound by silica gel column chromatography, eluting with 95:5DCM:NH₃ (7M in MeOH), affords the title compound (60 mg, 20%).

Step 2:4-{8-[3-Chloro-4-(4-methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5yl}-benzamide

In the same way as described for Compound 178, step 4, using(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-[3-chloro-4-(4-methyl-piperazin-1-yl)-phenyl]-amine(60.0 mg, 0.14 mmol),4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzamide (46.0 mg,0.28 mmol), Pd(PPh₃)₄ (40.0 mg, 0.035 mmol) and 1.5 M Na₂CO₃ (0.75 mL,1.12 mmol) in 2:1 DMF-dioxane (3 mL). The crude residue is trituratedwith DCM and then the title compound is crystallised from EtOH (4 mg,6%). LCMS: Rt 2.01 min (96%) m/z (APCI) 462 (M+H)⁺; ¹H-NMR (400 MHz,d₆-DMSO) δ (ppm). 2.27 (3H, s), 2.49-2.54 (4H, t), 2.99 (4H, m), 7.19(1H, d), 7.53 (1H, br s), 7.60 (1H, s), 7.76 (1H, s), 7.84 (2H, d), 7.98(1H, dd), 8.04 (1H, s), 8.09 (2H, d), 8.13 (1H, br s), 8.35 (1H, s),9.86 (1H, s).

Compound 139:5-{8-[4-(4-Methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-furan-2-carboxylic acid amide

In the same way as described for Compound 145, step 2 using[4-(4-methyl-piperazin-1-yl)-phenyl]-(5-tributylstannanyl-imidazo[1,2-a]pyrazin-8-yl)-carbamicacid tert-butyl ester (0.068 g, 0.162 mmol) in THF (3 mL),5-bromo-furan-2-carboxylic acid amide (0.061 g, 0.324 mmol) andPd(PPh₃)₄ (0.009 g, 0.008 mmol). The crude reaction mixture is stirredin (1:1) TFA:DCM (2 mL) for 2 hours at room temperature. Purification ofthe crude by silica gel column chromatography, eluting with DCM followedby 97:3 DCM:NH₃ (7M in MeOH), affords the title compound (7.4 mg, 11%).LCMS: Rt 1.81 min (99%) m/z (APCI) 418 (M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO)δ (ppm).2.26 (3H, s), 2.49-2.53 (4H, m), 3.14 (4H, m), 6.97 (2H, d),7.28 (1H, d, 7.34 (1H, d), 7.58 (1H, br s), 7.82 (1H, s), 7.90 (2H, d),8.02 (1H, br s), 8.15 (1H, s), 8.48 (1H, s), 9.62 (1H, s).

Compound 145:5-{8-[4-(4-Methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-thiophene-3-carboxylicacid amide

Step 1:[4-(4-Methyl-piperazin-1-yl)-phenyl]-(5-tributylstannanyl-imidazo[1,2-a]pyrazin-8yl)-amine

In the same way as described for Compound 212, step 2, using a solutionof(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-[4-(4-methyl-piperazin-1-yl)-phenyl]-carbamicacid tert-butyl ester (0.3 g, 0.62 mmol) in THF (5 mL), andisopropylmagnesium chloride (0.46 mL of a 2M solution in THF, 0.67mmol). After stirring for 5 min, tributyltin chloride (0.28 mL, 1.04mmol) is added and the reaction is stirred at −78° C. for 10 min beforebeing allowed to warm to room temperature. After stirring at roomtemperature for 30 min, the mixture is concentrated. The crude mixtureis purified by silica gel flash column chromatography using 95:5 DCM:NH₃(7M in MeOH), to give[4-(4-methyl-piperazin-1-yl)-phenyl]-(5-tributylstannanyl-imidazo[1,2-a]pyrazin-8-yl)-carbamicacid tert-butyl ester (0.32 g, 70%) as a yellow oil. LCMS: Rt 7.03 min(92.3%), m/z (ES⁺) 695, 697, 699.

Step 2:5-{8-[4-(4-Methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-thiophene-3-carboxylicacid amide

To a solution of[4-(4-methyl-piperazin-1-yl)-phenyl]-(5-tributylstannanyl-imidazo[1,2-a]pyrazin-8-yl)-carbamicacid tert-butyl ester (0.106 g, 0.15 mmol) in DMF (3 mL) is added5-bromo-thiophene-3-carboxylic acid amide (0.062 g, 0.3 mmol) andtetrakis(triphenylphosphine)palladium(0) (0.018 g, 0.015 mmol). Thesolution is degassed and heated at 85° C. for 18 hours. The reactionmixture is diluted with ethyl acetate and washed with water (3×) andbrine. The aqueous layers are backwashed with ethyl acetate and theorganic layers are combined, dried (MgSO₄), and concentrated. The crudemixture is then stirred in (1:1) TFA: dichloromethane (3 mL) for 2 hoursat room temperature. The resulting solution is diluted with ethylacetate and washed with water, sat. NaHCO₃ and brine. The aqueous layersare backwashed with ethyl acetate and the organic layers are combined,dried (MgSO₄), and concentrated. The crude material is purified by flashcolumn chromatography using 99:1 and 95:5 DCM:NH₃ (7M in MeOH), to give5-{8-[4-(4-methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-thiophene-3-carboxylicacid amide (0.013 g, 20%) as a yellow solid. LCMS: Rt 2.79 min, (93.3%),m/z (APCI) 434 (M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO) δ (ppm). 2.26 (3H, s),2.48-2.53 (4H, m), 3.14 (4H, m), 6.96 (2H, d), 7.42 (1H, br s), 7.61(1H, s), 7.78 (1H, s), 7.82 (2H, d), 7.92 (1H, s), 7.99 (1H, br s), 8.21(1H, s), 8.31 (1H, s), 9.61 (1H, s).

Compound 150:[4-(2-Morpholin-4-yl-ethoxy)-phenyl]-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine

Step 1: 4-(2-Morpholin-4-yl-ethoxy)-phenylamine

To a solution of 4-[2-(4-nitro-phenoxy)-ethyl]-morpholine (2.25 g, 8.93mmol) in ethanol (40 mL) is added palladium hydroxide (0.313 g, 0.45mmol) and the mixture is stirred in a Parr-apparatus under hydrogenpressure (5 bars) for 2 hours. Filtration over Celite 521 andevaporation gave a red oil purified by silica gel column chromatography.Elution with DCM and a mixture 90:10 DCM:MeOH affords the title compoundas a white solid (1.256 g, 63%). ¹H-NMR (400 MHz, CDCl₃) δ(ppm).2.52-2.63 (4H, m), 2.71-2.80 (2H, t), 3.46 (2H, br s), 3.70-3.78(4H, m), 4.00-4.10 (2H, m), 6.65 (2H, d), 6.77 (2H, d).

Step 2:(5-Bromo-imidazo[1,2-a]pyrazin-8-yl)-[4-(2-morpholin-4-yl-ethoxy)-phenyl]-amine

In the same way as described in the general procedure for aminedisplacement using 4-(2-morpholin-4-yl-ethoxy)-phenylamine (600 mg, 2.7mmol), N,N-diisopropylethylamine (0.470 mL, 2.7 mmol) and a solution of5,8-dibromo-imidazo[1,2-a]pyrazine (0.500 g, 1.80 mmol) in isopropanol(6 mL). Purification by silica gel column chromatography eluting with amixture 97:3 DCM:MeOH affords the title compound as a pale yellow solid(0650 mg, 86%). ¹H-NMR (400 MHz, CDCl₃) δ (ppm) 2.51-2.62 (4H, m),2.75-2.83 (2H, t), 3.71-3.78 (4H, m), 4.09-4.18 (2H, m), 6.90 (2H, d),7.51 (1H, s), 7.61 (1H, s), 7.64 (2H, d), 7.70 (1H, s), 7.90 (1H, s).LCMS: Rt 2.09 min (97%).

Step 3:[4-(2-Morpholin-4-yl-ethoxy)-phenyl]-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]amine

In the same way as described for Compound 178, step 4 using(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-[4-(2-morpholin-4-yl-ethoxy)-phenyl]-amine(113 mg, 0.27 mmol),4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (105 mg,0.541 mmol), 1.5M Na₂CO₃ (1.44 mL, 2.14 mmol), and Pd(PPh₃)₄ (0.78 g,0.67 mmol) in dioxane (4.3 mL). Purification by silica gel columnchromatography eluting with 96:4 DCM:NH₃ (7M in MeOH) followed bytrituration with diethyl ether, affords the title compound as a freebase (0.071 g, 65%). LCMS: Rt 1.59 min (100%), m/z (APCI) 406 (M+H)⁺;¹H-NMR (400 MHz, d₆-DMSO) δ (ppm). 2.40-2.60 (4H, m), 2.70-2.73 (2H, t),3.61-3.63 (4H, m), 4.10 (2H, m), 6.94-6.97 (2H, d), 7.58 (1H, s), 7.74(1H, s), 7.97 (2H, d), 8.06 (1H, s), 8.13 (1H, s), 8.41 (1H, s), 9.45(1H, s), 13.41 (1H, br s).

Compound 154:4-{8-[4-(4-Pyridin-2-yl-[1,2,3]triazol-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide

Step 1: 2-[1-(4-Nitro-phenyl)-1H-[1,2,3]triazol-4-yl]-pyridine

A mixture of 1-iodo-4-nitrobenzene (0.513 g, 2.06 mmol), 2-ethynylpyridine (0.208 mL, 2.06 mmol), L-proline (47.4 mg, 0.412 mmol), Na₂CO₃(43.7 mg, 0.412), NaN₃ (0.1607g, 2.47 mmol), (+)-sodium-L-ascorbate(40.8 mg, 0.206 mmol), copper (II) sulphate pentahydrate (16.4 mg, 0.103mmol) in 9:1 DMSO-water (4 mL) is stirred in a stem-tube at 65° C. for24 hours. After cooling, the reaction mixture is poured into ice-water(100 mL) and the resulting precipitate is collected by filtration,washed with 7% aq. NH₃ and dried in vacuo to give the title compound asa brown solid (0.323 g, 59%). HPLC (254 nm) Rt 2.36 min (89%).

Step 2: 4-(4-Pyridin-2-yl-[1,2,3]triazol-1-yl)-phenylamine

To a stirred solution of2-[1-(4-nitro-phenyl)-1H-[1,2,3]triazol-4-yl]-pyridine (0.149 g, 0.556mmol) in 1:1 THF/EtOH (8 mL) is added tin (II) dichloride dihydrate(0.439 g, 1.95 mmol) and the mixture is stirred at room temperature for22 hours. The solvent is removed in vacuo and 2N NaOH (3 mL) added tothe residue. The mixture is stirred for 1 hour, diluted with water andextracted with DCM (3×40 mL). The organic layers are dried over Na₂SO₄,filtered and evaporated to give a crude product purified by IsoluteFlashSilicaII Cartridge chromatography, eluting with 98:2 DCM:MeOH. Thefractions containing the title compound are combined and evaporated toafford the desired compound as a yellow solid (41.4 mg, 31%).

Step 3:(5-Bromo-imidazo[1,2-a]pyrazin-8-yl)-[4-(4-pyridin-2-yl-[1,2,3]triazol-1-yl)-phenyl]amine

In the same way as described for Compound 90, step 1, using5,8-dibromoimidazo[1,2-a]pyrazine (80.5 mg, 0.291 mmol),4-(4-pyridin-2-yl-[1,2,3]triazol-1-yl)-phenylamine (68.3 g, 0.288 mmol),NaO^(t)Bu (38.2 mg, 0.403 mmol), Pd₂(dba)₃ (10.5 mg, 0.0115 mmol),Xantphos (13.3 mg, 0.023mmol) and toluene (4 mL). The crude material ispurified by silica gel column chromatography eluting with 98:2 DCM:MeOHfollowed by 97:3 DCM:MeOH affording the title compound (52.8 mg, 42%).

Step 4:4-{8-[4-(4-Pyridin-2-yl-[1,2,3]triazol-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5yl}-benzamide

In the same way as described for Compound 178, step 4, using(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-[4-(4-pyridin-2-yl-[1,2,3]triazol-1-yl)-phenyl]-amine(40 mg, 0.0923 mmol), 4-(aminocarbonyl)phenylboronic acid (30.5 mg,0.185 mmol) 1.5 M Na₂CO₃ (0.492 mL, 0.738 mmol), and Pd(PPh₃)₄ (26.7 mg,0.023 mmol) in (2:1) DMF-dioxane (1.5 mL). The crude material ispurified by Isolute FlashSilicaII Cartridge chromatography eluting with98:2 DCM:NH₃ (7M in MeOH) followed by 97:3 DCM:NE₃ (7M in MeOH). Thetitle compound is obtained as a white solid (23.5 mg, 54%). LCMS: Rt2.67 min (95%) m/z (ES⁺) 474 (M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO) δ (ppm)7.41-7.47 (1H, d), 7.53 (1H, d), 7.66 (1H, br s), 7.81 (1H, s),7.84-7.90 (2H, d), 7.96-8.07 (3H, m), 8.08-8.20 (5H, m), 8.36-8.44 (2H,d), 8.70 (1H, d), 9.30 (1H, s), 10.15 (1H, s).

Compound 157:[4-(4-Methyl-piperazin-1-yl)-phenyl]-[5-(3-trifluoromethyl-1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine

Step 1: 4-Bromo-5-trifluoromethyl-pyrazole-1-carboxylic acid tert-butylester

To a solution of 4-bromo-5-trifluoromethyl-1H-pyrazole (0.515 g, 2.4mmol) in CH₃CN at 0° C is added (Boc)₂O (0.63 g, 2.89 mmol) followed byDMAP (0.29 g, 2.4 mmol). The reaction mixture is stirred overnight atroom temperature. After removing the solvent, the residue is dissolvedin DCM and washed with water, 2M HCl, sat. NaHCO₃ and brine. The organiclayer is dried over MgSO₄, filtered and concentrated in vacuo to affordthe title compound (0.55 g, 73%).

Step 2:[4-(4-Methyl-piperazin-1-yl)-phenyl]-[5-(3-trifluoromethyl-1H-pyrazol-4-yl)-imidazol[1,2-a]pyrazin-8-yl]-amine

In the same way as described for Compound 145, step 2 using[4-(4-methyl-piperazin-1-yl)-phenyl]-(5-tributylstannanyl-imidazo[1,2-a]pyrazin-8-yl)-carbamicacid tert-butyl ester (0.38 g, 0.78 mmol) in DMF (6mL),4-bromo-5-trifluoromethyl-pyrazole-1-carboxylic acid tert-butyl ester(0.49 g, 0.15 mmol) and Pd(PPh₃)₄ (0.09 g, 0.15 mmol). The crude mixtureis stirred in (1:1) TFA:DCM (2 mL) at room temperature for 3 hours.Purification by silica gel column chromatography eluting with DCMfollowed by a mixture of 99:1 and 95:5 DCM:NH₃ (7M in MeOH) affords thetitle compound (2.79 mg, 1%). LCMS: Rt 1.90 min (96%) m/z (ES⁺) 443(M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO) δ (ppm) 2.26 (3H, s), 2.49-2.54 (4H,m), 3.09-3.14 (4H, m), 6.96 (2H, d), 7.31 (1H, s), 7.63 (2H, d), 7.89(2H, d), 8.37 (1H, s), 9.44 (1H, s).

Compound 158:2-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-thiazole-5-carboxylicacid amide

Step 1:2-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-thiazole-5-carboxylicacid methyl ester

In the same way as described for Compound 212, step 3, using(4-morpholin-4-yl-phenyl)-(5-tributylstannanyl-imidazo[1,2-a]pyrazin-8-yl)-carbamicacid tert-butyl ester (0.25 g, 0.36 mmol), 2-bromo-thiazole-5-carboxylicacid methyl ester (0.159 g, 0.716 mmol), and Pd(PPh₃)₄ (0.041 g, 0.036mmol) in DMF (4 mL). The reaction mixture is concentrated, treated witha mixture of (1:1) TFA-DCM and stirred at room temperature for 18 hours.The mixture is diluted with EtOAc and washed with a solution of sat.NaHCO₃. The organic layers are dried over MgSO₄, filtered andconcentrated. Purification by silica gel column chromatography using75:25 petroleum ether-EtOAc affords the title compound (90 mg, 57%).HPLC (254 nm): Rt 3.89 min (73%)

Step 2:2-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-thiazole-5-carboxylacid amide

A solution of2-[8-(4-morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-thiazole-5-carboxylicacid methyl ester (90 mg, 0.21 mmol), 35% NH₄OH (2.5 mL), and NH₄Cl (0.2g), in MeOH (12 mL) is heated at 85° C. in a stem tube for 48 hours. Themixture is concentrated in vacuo and purified by silica gel columnchromatography eluting with 99:1 and 97:3 DCM:NH₃ (7M in MeOH).Trituration with diethyl ether-MeOH gives the title compound (4.9 mg,6%). LCMS: Rt 2.52 min (90%), m/z (ES⁺) 422 (M+H)⁺; ¹H-NMR (400 MHz,d₆-DMSO.) δ (ppm) 3.13 (4H, m), 3.79 (4H, m), 7.00 (2H, d), 7.75 (1H, brs), 7.81 (1H, s), 7.91 (2H, d), 8.25 (1H, s), 8.28 (1H, br s), 8.55 (1H,s), 9.08 (1H, s), 10.05 (1H, s).

Compound 159:2-Fluoro-4-{8-[4-(1-methyl-piperidin-4-ylmethoxy)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide

Step 1: 4-(1-Methyl-piperidin-4-ylmethoxy)-phenyl amine

In the same way as described for Compound 150, step 1, using1-methyl-4-(4-nitro-phenoxymethyl)-piperidine (0.65 g, 2.6 mmol) inethanol (40 mL) and palladium hydroxide (0.182 g, 0.26 mmol). Filtrationover Celite 521 and evaporation gave a red oil that was purified bysilica gel column chromatography eluting with DCM and a 80:20 mixtureDCM:MeOH. The title compound was obtained as a white solid (0.402 g,70%). ¹H-NMR (400 MHz, CDCl₃) δ (ppm) 1.30-1.51 (2H, m), 1.65-1.88 (3H,m), 1.89-2.09 (2H, t), 2.29 (3H, s), 2.90 (2H, d,), 3.45 (2H, br s),3.73 (2H, d), 6.67 (2H, d), 6.75 (2H, d).

Step 2:(5-Bromo-imidazo[1,2-a]pyrazin-8-yl)-[4-(1-methyl-piperidin-4-ylmethoxy)-phenyl]-amine

To a solution of 5,8-dibromo-imidazo[1,2-a]pyrazine (0.100 g, 0.36 mmol)in isopropanol (5 mL) are added 1,4-diazabicyclo[2,2,2]octane (DABCO)(0.036 mL, 0.36 mmol) and 4-(1-methyl-piperidin-4-ylmethoxy)-phenylamine(79.6 mg, 0.36 mmol). The reaction mixture is stirred at 90° C.overnight. The solvent is evaporated and the residue chromatographed onsilica gel column eluting with 97:3 followed by 94:6 and 90:10 DCM:MeOH.The title compound is obtained as a solid (20 mg, 13%). LCMS: Rt 2.30min(94%), m/z (APCI) 417.

Step 3:2-Fluoro-4-{8-[4-(1-methyl-piperidin-4-ylmethoxy)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide

In the same way as described for Compound 178, step 4, using(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-[4-(1-methyl-piperidin-4-ylmethoxy)-phenyl]-amine(20 mg, 0.048 mmol),2-fluoro-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzamide(25.5 mg, 0.10 mmol), Pd(PPh₃)₄ (13.9 mg, 0.001 mmol) and 1M Na₂CO₃ (0.8mL) in dioxane (2 mL). Purification by silica gel column chromatographyeluting with a mixture 95:5 followed by 90:10 DCM:MeOH affords a solidthat was triturated with ethyl acetate to give the title compound (19.4mg, 85%). LCMS: Rt 1.96min (94%), m/z (APCI) 475 (M+H)⁺; ¹H-NMR (400MHz, d₆-DMSO) δ (ppm) 1.48-1.61 (2H, m), 1.98-2.02 (3H, m), 2.51-2.58(4H, m), 2.70 (3H, br s), 3.91 (2H, d), 6.98 (2H, d), 7.55 (1H, s),7.65-7.87 (6H, m), 7.98 (2H, d), 8.06 (1H, s), 9.68 (1H, s).

Compound 160:[5-(5-Cyclopropyl-1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-(4-morpholin-4-yl-phenyl)-amine

Step 1: 4-Bromo-5-cyclopropyl-1H-pyrazole

To a solution of 2-cyclopropyl-1H-pyrazole (0.716 g, 6.62 mmol) in AcOH(10 mL) is added dropwise bromine (0.339 mL, 6.62 mmol). The reactionmixture is stirred vigorously for 10 min. The solvent is removed and theresidue partitioned between DCM and sat. Na₂CO₃. The aqueous phase (pH11) is extracted with DCM (3×). The organic layers are combined, driedover MgSO₄, filtered and concentrated to give a yellow solid (1.19 g,96%). The compound is used in the next step without furtherpurification.

Step 2:5-Cyclopropyl-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole

In the same way as described for Compound 209, step 2, using4-bromo-5-cyclopropyl-1H-pyrazole (0.477 g, 2.55 mmol),bis(pinacolato)diboron (1.29 g, 5.1 mmol), PdCl₂dppf (0.417 g, 0.51mmol) and KOAc (0.751 g, 7.65 mmol) suspended in DMSO (10 mL).Purification by silica gel column chromatography eluting with a 1:1mixture petroleum ether:EtOAc affords5-cyclopropyl-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole(214 mg, 14%). ¹H-NMR (400 MHz, CDCl₃) δ (ppm).0.83-1.05 (4H, m),1.25-1.42 (12H, m), 1.81 (1H, m), 7.78 (1H, s).

Step 3:5-(5-Cyclopropyl-1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-(4-morpholin-4-yl-phenyl)-amine

In the same way as described for Compound 178, step 4, using5-bromo-imidazo[1,2-a]pyrazin-8-yl)-4-morpholin-4-yl-phenylamine (80 mg,0.214 mmol),5-cyclopropyl-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole(219 mg, 0.37 mmol), Pd(PPh₃)₄ (49.4 mg, 0.04 mmol) and 0.5M Na₂CO₃(0.036 mL) in dioxane (2 mL). Purification by silica gel columnchromatography eluting with 97:3 DCM:MeOH, followed by crystallizationin DCM (drops of hexane) affords the title compound (13.3 mg, 15%).LCMS: Rt 2.12 min (98%) m/z (ES⁺) 402 (M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO)δ (ppm) 0.80-0.98 (4H, m), 1.70-1.98 (1H, m), 3.10 (4H, m), 3.79 (4H,m), 6.97 (2H, d), 7.40 (1H, s), 7.68 (1H, s), 7.77 (1H, s), 7.92-7.95(3H, m), 9.36 (1H, s), 12.65 (1H, d).

Compound 161:6Trifluoromethyl-4-[8-(4-morpholin-4-ylphenylamino)imidazo[1,2-a]pyrazin-5-yl]-1H-pyridin-2-one

Step 1: 2-Fluoro-3-iodo-6-(trifluoromethyl)pyridine

A solution of N,N-diisopropylamine (1.4 mL, 10.0 mmol) in anhydrous THF(15 mL) is cooled under N₂ to −78° C. and ^(n)BuLi (2.5 M in hexanes,4.0 mL, 10.0 mmol) is added dropwise at a rate to maintain thetemperature below −65° C. A solution of2-fluoro-6-(trifluoromethyl)pyridine (1.65 g, 10.0 mmol) in anhydrousTHF (15 mL) is added dropwise over 10 min and the resulting orangesolution is stirred for 2 h at −78° C. Iodine (2.54 g, 10.0 mmol) inanhydrous THF (12.5 mL) is then added dropwise over 15 min (keeping thetemperature under −65° C.), causing the colour of the solution to changeto red-brown and a precipitate to form. After 30 min the mixture isallowed to warm to 10° C. and the solvent is evaporated under reducedpressure. The residue is diluted with diethyl ether (25 mL) and washedwith Na₂S₂O₃ (2 M aq., 10 mL), HCl (2 M aq., 2×12.5 mL), NaHCO₃ (sat.aq., 12.5 mL) and brine (12.5 mL). The organic solvents are dried overNa₂SO₄ and evaporated under reduced pressure to afford a yellow oil. Theproduct is purified by silica chromatography, eluting with cyclohexane,to afford the title compound as a white solid (1.36 g). Alternatively,this compound may be used directly in the next step without furtherpurification.

Step 2: 2-Fluoro-4-iodo-6-(trifluoromethyl)pyridine

N,N-Diisopropylamine (3.0 mL, 21.0 mmol) is diluted with anhydrous THF(19 mL) and cooled to −30° C. ^(n)BuLi (2.5 M in hexanes, 8.4 mL, 21.0mmol) is added over 10 min, keeping the temperature under −20° C., andthe resulting solution is cooled to −78° C. A solution of2-fluoro-3-iodo-6-(trifluoromethyl)-pyridine (crude from previous step,2.82 g, theoretical 9.7 mmol) in anhydrous THF (4 mL) is added over 10min, keeping the temperature under −65° C., and the solution is stirredfor 5 min before HCl (2 M aq., 20 mL) is added and the mixture allowedto warm to rt. This is extracted with diethyl ether (3×35 mL) and thecombined solvents are dried over Na₂SO₄ and evaporated under reducedpressure to afford a brown oil. This is purified by silicachromatography, eluting with cyclohexane, to afford the title compoundas a pale yellow oil (2.33 g).

Step 3:2-Fluoro-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-6-(trifluoromethyl)pyridine

2-Fluoro-4-iodo-6-(trifluoromethyl)pyridine (145 mg, 0.50 mmol),bis(pinacolato)diboron (254 mg, 1.00 mmol), KOAc (147 mg, 1.50 mmol) andPd(dppf)Cl₂DCM (20 mg, 0.025 mmol) are stirred under N₂ in dioxane (0.5mL) and heated at 80° C. overnight. The mixture is cooled, diluted withNaHCO₃ (5% aq., 5 mL) and EtOAc (15 mL) and filtered through celite. Thelayers are separated and the aqueous phase extracted with further EtOAc(2×10 mL). The combined organic extracts are dried over Na₂SO₄ andevaporated under reduced pressure to afford the title compound as abrown oil which is used without further purification.

Step 4:[5-(2-Fluoro-6-trifluoromethyl-pyridin-4-yl)imidazo[1,2-a]pyrazin-8-yl]-(4-morpholin-4yl-phenyl)carbamicacid tert-butyl ester

(5-Bromoimidazo[1,2-a]pyrazin-8-yl)-(4-morpholin-4-ylphenyl)carbamicacid tert-butyl ester (50 mg, 0.11 mmol),2-fluoro-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-6-(trifluoromethyl)pyridine(126 mg, 0.21 mmol), KOAc (31 mg, 0.32 mmol) and Pd(dppf)Cl2DCM (4.3 mg,0.0053 mmol) are stirred under N₂ in dioxane (0.3 mL) and heated at 85°C. for 2.5 h. The mixture is cooled and partitioned between NaHCO₃ (5%aq., 5 mL) and EtOAc (15 mL). The layers are separated and the aqueousphase extracted with further EtOAc (2×10 mL). The combined organicextracts are washed with brine (10 mL), dried over Na₂SO₄ and evaporatedunder reduced pressure to afford the title compound as a brown oil (93mg) which was used without further purification.

Step 5:6-Trifluoromethyl-4-[8-(4-morpholin-4-ylphenylamino)imidazo[1,2-a]pyrazin-5-yl]-1H-pyridin-2-one

To a solution of[5-(2-fluoro-6-trifluoromethyl-pyridin-4-yl)imidazo[1,2-a]pyrazin-8-yl]-(4-morpholin-4-yl-phenyl)carbamicacid tert-butyl ester (92 mg, 0.12 mmol) in THF (0.5 mL) is added KOtBu(26 mg, 0.23 mmol) and the mixture is stirred at rt for 20 min. HCl (12M aq., 0.1 mL) is added and the mixture is diluted with water (10 mL)and concentrated under reduced pressure. The residue is dissolved in THF(5 mL) and HCl (12 M aq., 0.5 mL) is added and the mixture is stirred atrt for 5 h. NaHCO₃ (sat. aq.) is added until the pH is 7 and the mixtureis extracted with DCM (3×10 mL). The extracts are dried over Na₂SO₄ andevaporated under reduced pressure to afford a pale orange solid. This ispurified by preparative HPLC to afford the title compound as a yellowsolid (9.5 mg). LCMS: Rt=1.04 min (100%), m/z (ESI) 457 (M+H)⁺.

Compound 162:4-[3-Ethyl-8-(4-morpholin-4-ylphenylamino)-imidazo[1,2-a]pyrazin-5-yl]benzamide

This compound may be prepared using the methods as described forCompound 189, using 4-carbamoylbenzene boronic acid in Step 4. LCMS:Rt=1.03 min (100%), m/z (ESI) 443 (M+H)⁺.

Compound 166:4-{8-[4-(4-Dimethylaminomethyl-[1,2,3]triazol-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide

Step 1: Dimethyl-[1-(4-nitro-phenyl)-1H-[1,2,3]triazol-4-ylmethyl]-amine

In the same way as described for Compound 154, step 1 using1-iodo-4-nitrobenzene (0.498 g, 2.0 mmol), 1-dimethylamino-2-propyne(0.216 mL, 2.00 mmol), L-proline (46.1 mg, 0.40 mmol), Na₂CO₃ (42.4 mg,0.40), NaN₃ (0.156 g, 2.40 mmol), (+)-sodium-L-ascorbate (39.7 mg, 0.20mmol), copper(II) sulphate pentahydrate (16.0 mg, 0.10 mmol) in 9:1DMSO-water (4 mL). The residue is purified on Isolute FlashSilicaIIcartridge eluting with 97:3 DCM:NH₃ (7M in MeOH), to afford the titlecompound as a brown solid (0.179 g, 36%).

Step 2: 4-(4-Dimethylaminomethyl-[1,2,3]triazol-1-yl)-phenylamine

In the same way as described for Compound 154, step 2 usingdimethyl-[1-(4-nitro-phenyl)-1H-[1,2,3]triazol-4-ylmethyl]-amine (0.178g, 0.722 mmol), tin(II) chloride dehydrate (0.578 g, 2.54 mmol) in (1:1)THF/EtOH (4 mL). The residue is purified on Isolute FlashSilicaIIcartridge eluting with 95:5 DCM:NH₃ (7M in MeOH), to afford the desiredcompound as a yellow solid (93.2 mg, 59%).

Step 3:(5-Bromo-imidazo[1,2-a]pyrazin-8-yl)-[4-(4-dimethylaminomethyl-[1,2,3]triazol-1-yl)-phenyl]-amine

In the same way as described for Compound 90, step 1, using5,8-dibromoimidazo[1,2-a]pyrazine (116.9 mg, 0.422 mmol),4-(4-dimethylaminomethyl-[1,2,3]triazol-1-yl)-phenylamine (91.7 g, 0.422mmol), NaO^(t)Bu (56.8 mg, 0.591 mmol), Pd₂(dba)₃ (15.5 mg, 0.0169mmol), Xantphos (19.5 mg, 0.0337 mmol) and toluene (5 mL). The crudeproduct is purified by Isolute FlashSilicaII cartridge chromatographyeluting with 99:1 DCM/NH₃ (7M in MeOH), followed by 97:3 DCM:NH₃ (7M inMeOH). The title compound (63 mg, 36%) is isolated after a second columnchromatography purification eluting with 97:3 DCM:NH₃ (7M in MeOH).

Step 4:4-{8-[4-(4-Dimethylaminomethyl-[1,2,3]triazol-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide

In the same way as described for Compound 178, step 4, using(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-[4-(4-dimethylaminomethyl-[1,2,3]triazol-1-yl)-phenyl]-amine(63 mg, 0.152 mmol), 4-(aminocarbonyl)phenylboronic acid (50.3 mg, 0.305mmol) 1.5 M Na₂CO₃ (0.813 mL, 1.22 mmol) and Pd(PPh₃)₄ (44.0 mg, 0.0381mmol) in (2:1) DMF-dioxane (1.8 m1L). Purification of the crude materialby Isolute FlashSilicaII Cartridge chromatography eluting with 95:5DCM:NH₃ (7M in MeOH) followed by reverse phase preparative HPLC, affordsthe title compound as a white solid (4.7 mg, 7%). LCMS: Rt 1.92 min(96%) m/z (APCI) 455 (M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO) δ (ppm) 2.24 (6H,s), 3.61 (2H, s), 7.51 (1H, br s), 7.64 (1H, s), 7.80 (1H, s), 7.85-7.91(4H, m), 8.08-8.12 (3H), m), 8.16 (1H, br s), 8.36 (2H, d), 8.66 (1H,s), 10.09 (1H, s).

Compound 168:(4-Imidazol-1-yl-phenyl)-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine

Step 1:(5-Bromo-imidazo[1,2-a]pyrazin-8-yl)-(4-imidazol-1-yl-phenyl)-amine

In the same way as described for Compound 90, step 1, using5,8-dibromoimidazo[1,2-a]pyrazine (203.8 mg, 0.736 mmol),4-imidazol-1-yl-phenylamine (117.7 g, 0.739 mmol), NaO^(t)Bu (99 mg,1.03 mmol), Pd₂(dba)₃ (27.0 mg, 0.0295 mmol), Xantphos (34.1 mg, 0.0589mmol) and toluene (10 mL). The crude material is purified by IsoluteFlashSilicaII cartridge chromatography eluting with 99:1 DCM:NH₃ (7M inMeOH), followed by 97:3 DCM/NH₃ (7M in MeOH). The title compound isisolated, after a second column chromatography purification using a5:5:1 mixture EtOAc-petroleum ether-MeOH, as a yellow solid (104 mg,40%).

Step 2:(4-Imidazol-1-yl-phenyl)-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine

In the same way as described for Compound 178, step 4, using(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-(4-imidazol-1-yl-phenyl)-amine(49.8 mg, 0.14 mmol),4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole (54.4 mg,0.280 mmol), Pd(PPh₃)₄ (39.1 mg, 0.0338 mmol) and 1.5 M Na₂CO₃ (0.748mL, 1.12 mmol) in 2:1 DMF-dioxane (1.5 mL). Purification by IsoluteFlash SiII cartridge chromatography eluting with 98:2 DCM:NH₃ (7M inMeOH), followed by 97:3 DCM:NH₃ (7M in MeOH) and 95:5 DCM:NH₃ (7M inMeOH) affords the title compound as a white solid (26.5 mg, 55%). LCMS:Rt 1.75 min (96%), m/z (APCI) 343 (M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO) δ(ppm) 7.13 (1H, s), 7.63 (2H, d), 7.67 (1H, s), 7.73 (1H, s), 7.79 (1H,s), 8.09 (1H, br s), 8.18 (1H, s), 8.22 (1H, s), 8.27 (2H, d), 8.43 (1H,br s), 9.82 (1H, s), 13.39 (1H, br s).

Compound 169:[4-(2-Methyl-2H-tetrazol-5-yl)-phenyl]-[5-(1H-pyrazol-4-yl)-imidazol[1,2-a]pyrazin-8-yl]-amine

Step 1: 2-Methyl-5-(4-nitro-phenyl)-2H-tetrazole

To a stirred solution of 60% NaH (0.116 g, 2.89 mmol) and MeI (0.196 mL,3.15 mmol) in DMF (2 mL) (cooled to 0° C., under N₂) is added dropwise,a solution of 5-(4-nitrophenyl)-1H-tetrazole (0.502, 2.63 mmol) in DMF(7 mL). The reaction mixture is stirred under N₂ for 1 hour and 30 minat 5° C. and then quenched with water and extracted with EtOAc. Theorganic layers are combined and washed with sat. NaCl, dried over MgSO₄,filtered and evaporated to give a residue purified by silica gel columnchromatography. Elution with 80:20 followed by 60:40 petroleumether-EtOAc affords 2-methyl-5-(4-nitro-phenyl)-2H-tetrazole (0.265 g,49%) as a white solid and 1-methyl-5-(4-nitro-phenyl)-1H-tetrazole (37.7mg, 7%) as a yellow solid. The isomers are identified by NOEexperiments.

-   2-Methyl-5-(4-nitro-phenyl)-2H-tetrazole: ¹H-NMR (400 MHz, d₆-DMSO),    δ (ppm) 4.47 (3H, s) 8.34-8.40 (4H, m).-   1-Methyl-5-(4-nitro-phenyl)-1H-tetrazole: ¹H-NMR (400 MHz, d₆-DMSO)    δ (ppm) 4.25 (3H, s), 7.98-8.01 (2H, d), 8.44-8.47 (2H, d).    Step 2: 4-(2-Methyl-2H-tetrazol-5-yl)-phenylamine

A mixture of 2-methyl-5-(4-nitro-phenyl)-2H-tetrazole (0.260 g, 1.27mmol) and palladium on charcoal (53.6 mg) in ethanol (50 mL) is stirredin high pressure reactor under H2 at 4 bars for 17 hours. The catalystis removed by filtration through Celite 521, washing with EtOH. Thefiltrates are combined and concentrated in vacuo to afford the titlecompound as a white solid (0.213 g, 96%). HPLC (254 nm) 1.50 min (89%)

Step 3:(5-Bromo-imidazo[1,2-a]pyrazin-8-yl)-[4-(2-methyl-2H-tetrazol-5-yl)-phenyl]-amine

In the same way as described for Compound 90, step 1, using5,8-dibromoimidazo[1,2-a]pyrazine (334 mg, 1.21 mmol),4-(2-methyl-2H-tetrazol-5-yl)-phenylamine (211.1 g, 1.21 mmol),NaO^(t)Bu (162 mg, 1.69 mmol), Pd₂(dba)₃ (44.1 mg, 0.0482 mmol),Xantphos (55.8 mg, 0.0964 mmol) and toluene (16 mL). The crude productis purified by silica gel column chromatography eluting with a 8:2:1mixture petroleum ether:EtOAc:MeOH to afford the title compound as ayellow solid (198 mg, 44%).

Step 4:[4-(2-Methyl-2H-tetrazol-5-yl)-phenyl]-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8yl]-amine

In the same way as described for Compound 178, step 4, using(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-[4-(2-methyl-2H-tetrazol-5-yl)-phenyl]-amine(49.4 mg, 0.1333 mmol),4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole (51.6 mg,0.266 mmol), Pd(PPh₃)₄ (39.3 mg, 0.034 mmol), 1.5 M Na₂CO₃ (0.710 mL,1.07 mmol) in (2:1) DMF-dioxane (1.5 mL). Purification by IsoluteFlashSilicaII cartridge chromatography eluting with 97:3 DCM:NH₃ (7M inMeOH) affords the title compound as a white solid (17.3 mg, 36%). LCMS:Rt 2.60 min (96%), m/z 359 (M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO) δ (ppm)4.45 (3H, s), 7.72 (1H, s), 7.81 (1H, s), 8.05 (2H, d), 8.10 (1H, s),8.21 (1H, s), 8.33 (2H, d), 8.48 (1H, s), 9.91 (1H, s), 13.42 (1H, brs).

Compound 171:4-{8-[3-Fluoro-4-(4-methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide

Step 1:(5-Bromo-imidazo[1,2-a]pyrazin-8-yl)-[3-fluoro-4-(4-methyl-piperazin-1-yl)-phenyl]-amine

Following the general procedure for amine displacement using5,8-dibromoimidazo[1,2-a]pyrazine (0.50 g, 1.81 mmol),3-fluoro-4-(4-methyl-piperazin-1-yl)-phenylamine (0.45 g, 2.17 mmol) andDIPEA (0.47 mL, 2.72 mmol) in ^(i)PrOH (5 mL). The crude material ispurified by silica gel column chromatography eluting with 90:10 DCM:MeOHto give the title compound (35 mg, 5%).

Step 2:4-{8-[3-Fluoro-4-(4-methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide

In the same way as described for Compound 178, step 4, using(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-[3-fluoro-4-(4-methyl-piperazin-1-yl)-phenyl]-amine(60.0 mg, 0.15 mmol),4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzamide (49.5 mg,0.30 mmol), Pd(PPh₃)₄ (43.0 mg, 0.038 mmol) and 1.5 M Na₂CO₃ (0.80 mL,1.20 mmol) in dioxane (3 mL). The crude material is purified by silicagel column chromatography eluting with 95:5 and 90:10 DCM:MeOH to affordthe title compound (11 mg, 16%). LCMS: Rt 1.86 min (96%), m/z (ES⁺) 446(M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO) δ (ppm) 2.26 (3H, s), 2.52 (4H, m),3.01 (4H, m), 7.05 (1H, t), 7.52 (1H, br s), 7.59 (1H, s), 7.75 (1H, s),7.79-7.85 (3H, m), 8.04 (1H, s), 8.09-8.11 (3H, m), 8.15 (1H, br s),9.86 (1H, s).

Compound 173:5-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-thiazole-2-carboxylicacid amide

Step 1: 5-Bromo-thiazole-2-carboxylic acid amide

To 5-bromo-thiazole-2-carboxylic acid (0.267 g, 0.128 mmol) suspended indry dichloromethane (30 mL) are added oxalyl chloride (0.22 mL, 0.257mmol) and a drop of DMF. The reaction is stirred for 1 hour at roomtemperature. The solvent is evaporated and the resultant crude isdissolved in 7M NE₃ in MeOH. The reaction mixture is stirred overnightat room temperature. After evaporation of the solvent, the residue isdissolved in dichloromethane and washed with 1M NaOH. The organic phaseis dried over MgSO₄, filtered and evaporated. The residue is purified bysilica gel column chromatography eluting with 95:5 DCM:MeOH to give thetitle compound (33.8 mg, 25%).

Step 2:5-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-thiazole-2-carboxylicacid amide

In the same way as described for Compound 212, step 3, using(4-morpholin-4-yl-phenyl)-(5-tributylstannanyl-imidazo[1,2-a]pyrazin-8-yl)-carbamicacid tert-butyl ester (0.25 g, 0.365 mmol),5-bromo-thiazole-2-carboxylic acid amide (60.5 mg, 0.29 mmol) andPd(PPh₃)₄ (63.3 mg, 0.055 mmol) in DMF. Purification by silica gelcolumn chromatography eluting with 99:1 and 95:5 DCM:MeOH affords thetitle compound as a free base (27.4 mg, 49%).

Conversion into the mesylate salt following the usual procedure, using1M methanesulfonic acid (0.650 mL) affords the target compound (13.9 mg,41%). LCMS: Rt 2.57 min (97%), m/z (APCI) 422 (M+H)⁺; ¹H-NMR (400 MHz,d₆-DMSO) δ (ppm) 2.34 (3H, s, MsOH), 3.20 (4H, m), 3.82 (4H, t),6.99-7.09 (2H, m), 7.76 (1H, br s), 7.84 (1H, s), 7.94 (2H, d), 8.24(1H, s), 8.30 (1H, br s), 8.56 (1H, s), 9.09 (1H, s), 10.17 (1H, br s).

Compound 174:6-Methyl-4-[8-(4-morpholin-4-ylphenylamino)-imidazol[1,2-a]pyrazin-5-yl]-1H-pyridin-2-one

Step 1: 4-Bromo-6-methylpyridin-2-ol

A solution of 6-methylpyridine-2,4-diol (1.75 g, 14.0 mmol) in dry DMF(6.0 mL) is stirred under N₂ and phosphorus oxybromide (3.05 g, 10.6mmol) is added in one portion. The mixture is heated at 110° C. for 45min and then cooled and diluted with water (7 mL). Na₂CO₃ (s) is addedto bring the pH to 7 and the resulting suspension is cooled to 0° C. Theproduct is collected by suction filtration, rinsing with cold water anddiethyl ether, and dried overnight in a vacuum desiccator to afford thetitle compound as a light brown solid (920 mg).

Step 2: 4-Bromo-2-(tert-butyldimethylsilyloxy)-6-methylpyridine

A solution of 4-bromo-6-methylpyridin-2-ol (376 mg, 2 mmol) in dry DMF(20 mL) is stirred under N₂. TBDMS-Cl (600 mg, 4 mmol) is added,followed by triethylamine (0.97 mL, 7.00 mmol), and the mixture isstirred at rt for 1 h. The solution is poured into ice-water (170 mL)and extracted with diethyl ether (5×50 mL). The combined extracts aredried over Na₂SO₄ and the solvent is evaporated under reduced pressure.The residue is taken up in pentane (50 mL), dried over Na₂SO₄ and thesolvent evaporated again, to afford the crude title compound as a yellowoil. This is used without further purification.

Step 3:(4-Morpholin-4-ylphenyl)-[5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)imidazo[1,2-a]pyrazin-8-yl]carbamic acid tert-butyl ester

(5-Bromoimidazo[1,2-a]pyrazin-8-yl)-(4-morpholin-4-ylphenyl)carbamicacid tert-butyl ester (47 mg, 0.10 mmol), bis(pinacolato)diboron (51 mg,0.20 mmol), KOAc (29 mg, 0.30 mmol) and Pd(dppf)Cl₂-DCM (4 mg, 0.0050mmol) are stirred under N₂ in dioxane (0.3 mL) and heated at 80° C. for1.5 h. The resulting solution is cooled and used for the following step.

Step 4:6-Methyl-4-[8-(4-morpholin-4-ylphenylamino)-imidazo[1,2-a]pyrazin-5-yl]-1H-pyridin-2-one

Half of the solution prepared in Step 3 and4-bromo-2-(tert-butyldimethylsilyloxy)-6-methylpyridine (30 mg, 0.10mmol) are mixed and diluted with dioxane (0.15 mL). KOAc (15 mg, 0.15mmol) and Pd(dppf)Cl₂DCM (2 mg, 0.0025 mmol) are added and the system isflushed with N₂ and heated at 80° C. for 2.5 h. The mixture is cooled tort, diluted with DCM (5 mL) and MeOH (1 mL) and filtered through celite.The solvents are evaporated, the residue is suspended in DCM (2 mL) andTFA (0.5 mL) is added, causing the solid to dissolve. The solution isstirred at rt overnight and the solvents are then evaporated. Theresidue is purified by preparative HPLC to afford the title compound asa yellow solid. LCMS: Rt=0.84 min (100%), m/z (ESI) 403 (M+H)⁺.

Compound 175:4-[8-(3-Carbamoyl-4-morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-2,6-difluoro-benzamide

Step 1:5-(5-Bromo-imidazo[1,2-a]pyrazin-8-ylamino)-2-morpholin-4-yl-benzamide

In the same way as described in the general procedure for aminedisplacement using 5,8-dibromo-imidazo[1,2-a]pyrazine (0.131 g, 0.47mmol), 5-amino-2-morpholin-4-yl benzamide (157 mg, 0.71 mmol),N,N-diisopropylethylamine (0.120 mL, 0.71 mmol) in iso-propanol (3.7mL). Trituration with ^(t)PrOH and Et₂O affords the title compound (127mg, 67%).

Step 2: 4-Bromo-2,6-difluoro-benzamide

To a solution of 4-bromo-2,6-difluoro-benzoic acid (2.77 g, 11.7 mmol)and 1-hydroxybenzotriazole (1.74 g, 12.8 mmol) is added1-ethyl-3-(3′-dimethylaminopropyl)carbodiimide (2.46 g, 12.9 mmol) andthe solution is stirred at room temperature for 2 hours. After coolingat 0° C. is added aq. NH3 (1.74 mL) and the reaction mixture is stirredfor an additional 20 hours. The solvent is evaporated in vacuo and theresidue partitioned between diethyl ether and water. The aqueous phaseis extracted several times with Et₂O, the organic layers are thencombined, washed with 1M HCl, sat. NaHCO₃, dried over MgSO₄, filteredand concentrated in vacuo to afford the benzamide as a white solid (2.56g, 93%).

Step 3: 2,6-Difluoro-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzamide

In the same way as described for Compound 209, step 2 using4-bromo-2,6-difluoro-benzamide (2.50 g, 10.6 mmol),bis(pinacolato)diboron (2.96 g, 11.7 mmol), PdCl₂dppf (0.260 g, 0.32mmol) and KOAc (3.12 g, 31.6 mmol) suspended in dioxane (30 mL). Thereaction mixture is filtered through silica and washed with DCM. Afterevaporation of the solvent, the residue is dissolved in EtOAc and washedwith water and brine. The organic layer is dried over MgSO₄, filteredand concentrated in vacuo to afford the boronic ester as a brown solid(2.95 g, 98%).

Step 4:4-[8-(3-Carbamoyl-4-morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-2,6-difluoro-benzamide

In the same way as described for Compound 178, step 4, using5-(5-bromo-imidazo[1,2-a]pyrazin-8-ylamino)-2-morpholin-4-yl-benzamide(120 mg, 0.29 mmol),2,6-difluoro-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzamide(163 mg, 0.58 mmol) 1.5M K₂CO₃ (1.7 mL, 2.55 mmol), and Pd(PPh₃)₄ (84mg, 0.073 mmol) in dioxane (3 mL). Purification by silica gel columnchromatography eluting with 98:8 DCM:NH₃ (7M in MeOH) affords the titlecompound as a pale yellow solid (30 mg, 21%). Following the usualprocedure, using methanesulfonic acid (0.593 mL), the free base (29.3mg, 0.060 mmol) is converted into the mesylate salt (27 mg, 99%). LCMS:Rt 2.47 min (98%), m/z (ES⁺) 494 (M+H)⁺, ¹H-NMR (400 MHz, d₆-DMSO) δ(ppm.) 2.35 (3H, s, MsOH), 2.97-3.05 (4H, m), 3.74-3.83 (4H, t), 7.35(1H, d), 7.60-7.65 (4H, m), 7.80 (1H, s), 7.98 (1H, br s), 8.05-8.08(1H, m), 8.13 (1H, s), 8.26 (1H, br s), 8.50 (1H, d), 8.68 (1H, br s),10.00 (1H, s).

Compound 177:4-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-thiophene-2-carboxylicacid amide

Step 1: 4-Bromo-thiophene-2-carboxylic acid amide

In the same way as described for Compound 179, step 2, using 4bromo-thiophene-2-carboxylic acid (2.0 g, 9.66 mmol),1-ethyl-3-(3′-dimethylaminopropyl)carbodiimide hydrochloride (2.04 g,10.63 mmol), 1-hydroxybenzotriazole hydrate (1.44 g, 10.63 mmol) and aq.NH₃ (1 ml, 17.3 mmol) in DMF (20 ml). Water is added to the reactionmixture and the resultant precipitate is collected by filtration andwashed with 1M NaOH, H₂O and petrol. The title compound is isolated as awhite solid (1.56 g, 78%).

Step 2:4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-thiophene-2-carboxylicacid amide

In the same way as described for Compound 209, step 2 using4-bromo-thiophene-2-carboxylic acid amide (1.3 g, 6.34 mmol),bis(pinacolato)diboron (3.22 g, 12.68 mmol), PdCl₂dppf (0.26 g, 0.318mol) and KOAc (1.87 g, 19.10 mmol) in dioxane (20 mL). The title productis crystallised from EtOAc-petroleum ether (2.135 g, 77% pure by LCMS).

Step 3:4-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-thiophene-2-carboxylicacid amide

In the same way as described for Compound 178, step 4, using5-bromo-imidazo[1,2-a]pyrazin-8-yl)-4-morpholin-4-yl-phenylamine (100mg, 0.268 mmol),4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-thiophene-2-carboxylicacid amide (136 mg, 0.536 mmol), Na₂CO₃ (85.3 mg, 0.804 mmol), andPd(PPh₃)₄ (77.5 mg, 0.067 mmol) in dioxane:water (2.5 mL). The crudecompound is precipitated from THF, filtered, dissolved in MeOH andconverted into the mesylate salt using the procedure describedpreviously employing methanesulfonic acid (1.08 mL, 0.108 mmol).Trituration with diethyl ether gives the mesylate salt as a yellow solid(48.8 mg, 43%). LCMS: Rt 2.19 min (96%), m/z (APCI) 421 (M+H)⁺; ¹H-NMR(400 MHz, d₆-DMSO) δ (ppm) 2.35 (3H, s, MsOH), 3.10-3.19 (4H, m), 3.79(4H, t), 7.05 (2H, d), 7.51 (1H, s), 7.59 (1H, br s), 7.82-7.84 (3H, m),8.16 (3H, m), 8.21 (1H, s), 9.93 (1H, br s).

Compound 178:5-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-2,3-dihydro-isoindol-1-one

Step 1: 4-Bromo-2-bromomethyl-benzoic acid methyl ester

4-Bromo-2-methyl-benzoic acid (4.6 g, 21.39 mmol) is dissolved in 2M HClin MeOH and refluxed for 3 hours. The solvent is evaporated to give the4-bromo-2-methyl-benzoic acid methyl ester (4.24 g, 86%). Thisintermediate (18.51 mmol) is dissolved in carbon tetrachloride (100 mL)and N-bromosuccinimide (NBS) (5.57 g, 24.06 mmol) is added. AIBN (122mg, 740 □mol) is then added and the mixture purged with nitrogen for 5min. The reaction mixture is then refluxed for 4 hours. After cooling toroom temperature the reaction mixture is filtered and the filtrate isevaporated. The residue is purified by flash chromatography (silica gel,2:1 petroleum ether/ethyl acetate) to give the title compound (3.42 g,60%).

Step 2: 5-Bromo-2,3-dihydro-isoindol-1-one

4-Bromo-2-bromomethyl-benzoic acid methyl ester (0.5 g, 16.2 mmol) istreated with methanolic ammonia (10 mL, 7 N NH₃ in MeOH) for 5 minutesat 90° C. After cooling to room temperature a precipitate is formed,collected by filtration and washed with a small amount of methanol toafford the title compound as a colourless solid (224 mg, 65%). ¹H-NMR(400 MHz, d₆-DMSO) δ (ppm) 4.41 (2H, s), 7.64 (1H, d), 7.70 (1H, d),7.87 (1H, s), 8.67 (1H, br s). LCMS: Rt 2.49 min, (99.6%), m/z (APCI)212 (M+H⁺).

Step 3:5-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-2,3-dihydro-isoindol-1-one

5-Bromo-2,3-dihydro-isoindol-1-one (230 mg, 1.08 mmol),bis(pinacolato)diboron (300 mg, 1.18 mmol), PdCl₂dppf (25 mg, 31□mol)and KOAc (320 mg, 3.26 mmol) are suspended in dioxane (4 mL), purgedwith nitrogen for 5 minutes and then heated at 85° C. overnight. Thesolvent is removed in vacuo and the residue partitioned between ethylacetate and water. The aqueous layer is extracted with ethyl acetate(3×) and the combined organic phases are washed once with brine,filtered through MgSO₄ and evaporated. The solid residue is trituratedwith hexane and dried in vacuo to yield the title compound (185 mg, 66%)as a grey solid. ¹H-NMR (400 MHz, CDCl₃) δ (ppm) 1.37 (12H, s), 4.45(2H, s), 6.38 (1H, br s), 7.87 (1H, d), 7.93 (2H, m).

Step 4:5-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-2,3-dihydro-isoindol-1-one

A degassed solution of5-bromo-imidazo[1,2-a]pyrazin-8-yl)-4-morpholin-4-yl-phenylamine (1.0 g,2.06 mmol),5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-2,3-dihydro-isoindol-1-one(1.04 g, 4.02 mmol), 1.5 M Na₂CO₃ (14.3 mL, 21.44 mmol) and Pd(PPh₃)₄(0.77 g, 0.67 mmol) in dioxane (40 mL) is stirred overnight at 90° C.The solvent is removed in vacuo and the residue partitioned betweenethyl acetate and water. A solid is formed and collected by filtration.The aqueous layer is extracted with ethyl acetate (2×) and the combinedorganic phases and the solid are dissolved using MeOH/DCM and thenevaporated to dryness. The residue is purified by silica gel columnchromatography eluting with DCM followed by 95:5 DCM:NH₃ (7M in MeOH).The fractions containing the desired product are combined and evaporatedto afford the title compound as a solid.

After trituration with diethyl ether, the compound (0.93 g, 2.18 mmol,83%) is converted into the mesylate salt. The solid is dissolved in theminimum amount of MeOH/DCM, (refluxed to dissolve) and 1M methanesulfonic acid in MeOH (2.18 mL, 2.18 mmol) is added. The mixture iscooled and concentrated to afford a solid that is triturated severaltimes with a mixture of 1:1 ethyl acetate-diethyl ether and DCM-diethylether, filtered and dried in vacuo to afford the desired compound as ayellow solid (0.954 g, 84%). LCMS: Rt 2.32 min (98.4%); m/z (APCI) 427(M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO) δ (ppm) 2.39 (3H, s, MsOH), 3.29 (4H,m), 3.85 (4H, m), 4.52 (2H, s), 7.22 (2H, d), 7.45 (1H, s), 7.68-7.82(3H, m), 7.89-7.93 (3H, m), 8.14 (1H, s), 8.77 (1H, s), 10.77(1H, br s).

Compound 179:5-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-furan-3-carboxylicacid amide

Step 1: 5-Bromo-furan-3-carboxylic acid

Furan-3-carboxylic acid (1 g, 8.93 mmol) is added to a solution ofpyridinium hydrobromide perbromide (3.02 g, 9.46 mmol) in acetic acid (5mL). The reaction mixture is stirred at 50° C. for 5 hours, then at roomtemperature overnight. After removing the solvent in vacuo, water isadded and a precipitate is formed. The solid is collected by filtrationand dried to yield the title compound (0.364 g, 21%). HPLC (254 nm): Rt2.65 min (88%).

Step 2: 5-Bromo-furan-3-carboxylic acid amide

A solution of 5-bromo-furan-3-carboxylic acid (0.364 g, 1.92 mmol),3-hydroxybenzotriazole hydrate (0.28 g, 2.11 mmol),1-ethyl-3-(3′-dimethylaminopropyl)carbodiimide (0.40 g, 2.11 mmol) inDMF (11 mL) is stirred at room temperature for 2 hours. The reactionmixture is then cooled to 0° C. and aq. NH₃ (0.22 mL) is added. Themixture is stirred at room temperature for an additional 5 hours, thenthe solvent is removed in vacuo and the residue dissolved in EtOAc,washed with 1N NaOH, and 1N HCl. The organic layers are combined, driedover MgSO₄ and concentrated to afford the title compound (0.288 g, 79%).HPLC (254 nm): Rt 2.17 min (85.3%).

Step 3:5-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-furan-3-carboxylicacid amide

In the same way as described for Compound 212, step 3, using(4-morpholin-4-yl-phenyl)-(5-tributylstannanyl-imidazo[1,2-a]pyrazin-8-yl)-carbamicacid tert-butyl ester (0.25 g, 0.365 mmol), 5-bromo-furan-3-carboxamide(0.139 g, 0.66 mmol) and Pd(PPh₃)₄ (63.3 mg, 0.05 mmol) in DMF (10 mL).Purification by silica gel column chromatography eluting with DCMfollowed by 97:3 and 94:6 DCM:MeOH gives(4-morpholin-4-yl-phenyl)-(5-tributylstannanyl-imidazo[1,2-a]pyrazin-8-yl)-carbamicacid tert-butyl ester as a yellow solid (153 mg, 83%). HPLC (254 nm): Rt2.72 min (93.7%), m/z 505 (M+H)⁺

The solid is dissolved in dioxane:4M HCl (4 mL). A few drops of methanolare added to achieve complete dissolution. The yellow solution isstirred at room temperature for 20 min. A precipitate is formed andcollected by filtration, washed with diethyl ether and dried in vacuo toafford5-[8-(4-morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-furan-3-carboxylicacid amide as the hydrochloride salt (111.6 mg, 91%). HPLC (254 nm) (HClsalt): Rt 2.31 min (98.2%), m/z 405 (M+H)⁺. The hydrochloride salt isconverted into the free base suspending in ethyl acetate and washingwith 1M Na₂CO₃.

Formation of the mesylate salt, following the procedure describedpreviously, using5-[8-(4-morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-furan-3-carboxylicacid amide (83 mg, 0.205 mmol) and methanesulfonic acid (2.06 mL)affords the title compound as a solid (95.4 mg, 93%). LCMS: Rt 2.34 min(96.2%), m/z (APCI) 405 (M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO) δ (ppm) 2.35(3H, s, MsOH), 3.28 (4H, m), 3.86 (4H, m), 7.21 (2H, br d), 7.42 (1H, brs), 7.50 (1H, s), 7.80-8.07 (5H, m), 8.40 (1H, s), 8.49 (1H, s), 10.34(1H, br s).

Compound 183:N-{5-[5-(1H-Pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-ylamino]pyridin-2-yl}benzamide

This compound may be prepared using the same methods as described forcompound 200, using benzoyl chloride in Step 1. LCMS: Rt=1.13 min(100%), m/z (ESI) 397 (M+H)⁺.

Compound 184:2-Methoxy-N-(6-methylpyridin-3-ylmethyl)-4-[5-(1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-ylamino]benzamide

Step 1: 2-Methoxy-N-(6-methylpyridin-3-yl)methyl-4-nitrobenzamide

2-Methoxy-4-nitrobenzoic acid (293 mg, 1.49 mmol) is dissolved in DMF (2mL) and 4-methylmorpholine (220 μL, 3.0 mmol) and TBTU (1.79 g, 1.7mmol) are added. The mixture is stirred at rt for 30 min andC-(6-methylpyridin-3-yl)methylamine (400 mg, 3.27 mmol) is added.Stirring is continued at rt for 12 h. DCM (10 mL) is added and theorganic phase is washed with Na₂CO₃ (5% aq.), HCl (3% aq.) and water,and then dried over Na₂SO₄. After evaporation of the solvents, theresidue is triturated with ether-hexane to afford the title compound asa white solid.

Step 2: 4-Amino-2-methoxy-N-[(6-methylpyridin-3-yl)methyl]benzamide

A solution of 2-methoxy-N-(6-methylpyridin-3-yl)methyl-4-nitrobenzamide(448 mg, 1.49 mmol) in EtOH and EtOAc (8 mL each) is stirred andammonium formate (375 mg, 6 mmol) and 10% Pd/C (100 mg) are added. Themixture is heated at reflux for 20 min, cooled, filtered through celite,the solid is washed with EtOH and the combined solvents are evaporatedto afford the title compound.

Step 3:4-(5-Bromo-imidazo[1,2-a]pyrazin-8-ylamino)-2-methoxy-N-(6-methylpyridin-3-ylmethyl)benzamide

5,8-dibromoimidazo[1,2-a]pyrazine (285 mg, 1.03 mmol) and4-amino-2-methoxy-N-[(6-methylpyridin-3-yl)methyl]benzamide (280 mg,1.03 mmol) are stirred in ^(i)PrOH (5 mL) and HBr (48% aq., 380 μL) isadded. The mixture is heated at reflux for 24 hours. The cooledsuspension is poured into NaHCO₃ (sat. aq., 25 mL) and water (25 mL) andextracted with CDCl₃ (3×40 mL). The extracts are dried over MgSO₄ andevaporated. The residue is purified by column chromatography, elutingwith 10% -20% DCM/MeOH to afford the title compound as a pale yellowsolid (200 mg, 0.43 mmol).

Step 4:2-Methoxy-N-(6-methylpyridin-3-ylmethyl)-4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino-benzamide

Pyrazole-4-boronic acid (19 mg, 0.17 mmol),4-(5-bromo-imidazo[1,2-a]pyrazin-8-ylamino)-2-methoxy-N-(6-methylpyridin-3-ylmethyl)benzamide(40 mg, 0.085 mmol), K₂CO₃ (24 mg, 0.17 mmol) and Pd(dppf)Cl₂-CH₂Cl₂ (4mg, 0.005 mmol) are weighed into a sealable tube. The tube is flushedwith nitrogen and dioxane-water (4:1, 4 mL) is added. The tube issealed, placed in an ultrasonic bath under a flow of nitrogen gas for 30seconds and then placed into an oil bath at 85° C. The reaction isstirred for 28 hours, adding additional portions of boronic acid (10 mg)and catalyst (2 mg) after 2 h and 18 h. The crude mixture is absorbedonto SiO₂ and purified by column chromatography, eluting with 5% MeOH inDCM and then 5% 2 M NH₃/MeOH in DCM to afford the title compound as abrown powdery solid (27.8 mg). ¹H NMR (400 MHz, DMSO-d₆): δ=13.40 (1H,br s); 9.83 (1H, s); 8.62 (1H, t, J=5 Hz); 8.44-8.42 (2H, m); 8.18 (1H,s); 8.08 (1H, br s); 8.02 (1H, d, J=2 Hz); 7.91 (1H, dd, J=8 Hz, 2 Hz);7.81 (1H, d, J=9 Hz); 7.78 (1H, s); 7.69 (1H, s); 7.62 (1H, dd, J=8 Hz,2 Hz); 7.21 (1H, d, J=8 Hz); 4.48 (2H, d, J=6 Hz); 3.93 (3H, s); 2.44(3H, s). LCMS: Rt 0.83 min (100%) m/z (ESI) 455 (M+H)⁺.

Compound 189:[3-Ethyl-5-(1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-yl]-(4-morpholin-4-yl-phenyl)amine

This compound may be prepared using the methods described for Compound131, using butanal in step 1. LCMS: Rt=0.94 min (100%), m/z (ESI) 390(M+H)⁺.

Compound 190:4-(8-{3-Methoxy-4-[(6-methylpyridin-3-ylmethyl)carbamoyl]phenylamino}imidazo[1,2-a]pyrazin-5-yl)thiophene-2-carboxylic acid amide

This compound may be prepared using the method described for compound184, using 2-(aminocarbonyl)thiophene-4-boronic acid in Step 4. LCMS:Rt=0.95 min (100%), m/z (ESI) 514 (M+H)⁺.

Compound 192:4-(8-{4-[1-(2,2,2-Trifluoroethyl)piperidin-4-yl]phenylamino}imidazo[1,2-a]pyrazin-5-yl)thiophene-2-carboxylicacid amide

Step 1: 1-Trifluoroacetyl-4-(4-nitrophenyl)piperidine

Triethlyamine (1.0 mL, 7.3 mmol) and 4-(4-nitrophenyl)piperidine (1.0 g,4.8 mmol) are stirred in DCM (25 mL) at 0° C. under N₂ andtrifluoroacetic anhydride (0.81 mL, 5.8 mmol) is added. The mixture isstirred for three days, allowing the temperature to warm to rt. Thesolution is then diluted with DCM (50 mL) and washed with water (2×15mL), NaHCO₃ (50% sat. aq., 2×15 mL) and brine (15 The solvent is driedover MgSO₄ and evaporated to afford the desired compound (1.46 g, 4.66mmol).

Step 2: 1-(2, 2, 2-Trifluoroethyl)-4-(4-nitrophenyl)piperidine

A solution of 1-trifluoroacetyl-4-(4-nitrophenyl)piperidine (1.42 g, 4.7mmol) in THF (15 mL) is stirred in 25 mL 2-necked flask fitted with acondenser and pressure-equalising addition funnel. The system is flushedwith N₂, NaBH₄ (210 mg, 5.6 mmol) is added and the flask is cooled to 0°C. A solution of iodine (600 mg, 2.3 mmol) in THF (5 mL) is then addeddropwise over 20 minutes, after which the addition funnel is removed andthe mixture heated at reflux overnight. The resulting pale yellowsuspension is cooled to rt and MeOH (1.5 mL) is added cautiously,causing vigorous evolution of a gas. Evaporation of the solvents affordsthe title compound, which is used without further purification.

Step 3: 4-[1-(2,2,2-Trifluoroethyl)piperidin-4-yl]phenylamine

Ammonium formate (1.38 g, 22 mmol) and 10% Pd/C (230 mg, 0.2 mmol) areadded to a solution of1-(2,2,2-trifluoroethyl)-4-(4-nitrophenyl)piperidine (1.26 g, 4.4 mmol)in EtOH (10 mL) and EtOAc (10 mL). The suspension is heated at refluxfor 24 hours, adding further portions of ammonium formate (2 g) after 4h and 8 h. The mixture is filtered through celite and evaporated toafford an orange solid. This is partitioned between DCM (40 mL) andwater (20 mL) and the layers separated. The aqueous phase is extractedwith DCM (2×20 mL) and the combined organic layers are dried over MgSO4and evaporated under reduced pressure to affordN-{4-[l-(2,2,2-trifluoroethyl)piperidin-4-yl]phenyl}formamide as a paleorange solid (980 mg).

A solution of the formamide in MeOH (20 mL) is stirred at rt and HCl(conc., 1 mL) is added. The deep purple solution is heated at reflux for1 h, cooled and the MeOH is evaporated. The residue is stirred withwater (20 mL) and NaHCO₃ (sat. aq.) is added until bubbling ceases. Themixture is extracted with DCM (20 mL, 2×10 mL) and the combined extractsare dried over MgSO₄ and evaporated under reduced pressure to afford thetitle compound as an orange solid (870 mg).

Steps 4 and 5:4-(8-{4-[1-(2,2,2-Trifluoroethyl)piperidin-4-yl]phenylamino}imidazo[1,2-a]pyrazin-5-yl)thiophene-2-carboxylicacid amide

These steps may be performed using the methods as described for Compound202, step 3 and for Compound 200, Step 4. LCMS: Rt=2.10 min (100%), m/z(ESI) 501 (M+H)⁺.

Compound 198:2-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-4,5-dihydro-thieno[2,3-c]pyrrol-6-one

Step 1: 3-Bromomethyl-thiophene-2-carboxylic acid methyl ester

3-Methyl-thiophene-2-carboxylic acid methyl ester (1 g, 6.4 mmol) isdissolved in CCl₄ (15 mL), and NBS (1.19 g, 6.7 mmol) and benzoylperoxide (15 mg) are added. The reaction mixture is refluxed for 6hours, then water is added and the mixture is extracted with EtOAc (3×).The organic layers are dried over MgSO₄, filtered and concentrated. Thecrude product is purified by silica gel column chromatography elutingwith 98:2 petroleum ether-EtOAc to give the title compound (1.34 g,27%).

Step 2: 3-Aminomethyl-thiophene-2-carboxylic acid methyl ester

3-Bromomethyl-thiophene-2-carboxylic acid methyl ester (1.297 g, 5.54mmol) is dissolved in DMF (30 mL) and MeOH/NH₃ is added (30 mL). Thereaction mixture is stirred at room temperature for 1 hour, andconcentrated in vacuo. Purification of the residue by silica gel columnchromatography eluting with 95:5 and 93:7 DCM:MeOH affords the titlecompound (0.82 g, 86%).

Step 3: 4,5-Dihydro-thieno[2,3-c]pyrrol-6-one

To a solution of 3-aminomethyl-thiophene-2-carboxylic acid methyl ester(0.82 g, 4.79 mmol) in 1:1 MeOH-EtOH (125 mL), is added K₂CO₃ (0.66 g,4.79 mmol) and the mixture is stirred at 90° C. for 5 hours. The solventis removed in vacuo and the residue purified by silica gel columnchromatography eluting with 97:3 DCM:MeOH. The title compound isisolated (0.365 g, 55%).

Step 4: 2-Bromo-4,5-dihydro-thieno[2,3-c]pyrrol-6-one

4,5-Dihydro-thieno[2,3-c]pyrrol-6-one (0.365 g, 2.63 mmol) is dissolvedin acetonitrile (10 mL), and NBS (0.47 g, 2.63 mmol) is slowly added at−10° C. The reaction mixture is stirred at room temperature overnight.The solvent is removed in vacuo and the residue taken-up in EtOAc andwashed with 1M NaOH. The organic phase is dried over MgSO₄, filtered andconcentrated in vacuo. The crude is purified by silica gel columnchromatography eluting with a 50:50 mixture and then 50:70 petroleumether-EtOAc. The title compound is isolated (91.1 mg, 16%). HPLC (254nm): Rt 2.55 min (70.3%), m/z 218/220 (M+H)⁺.

Step 5: 2-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2,-a]pyrazin-5-yl]-4,5-dihydro-thieno[2,3-c]pyrrol-6-one

In the same way as described for Compound 212, step 3, using(4-morpholin-4-yl-phenyl)-(5-tributylstannanyl-imidazo[1,2-a]pyrazin-8-yl)-carbamicacid tert-butyl ester (0.28 g, 0.41 mmol),2-bromo-4,5-dihydro-thieno[2,3-c]pyrrol-6-one (0.06 g, 0.276 mmol), andPd(PPh₃)₄ (0.032 g, 0.0276 mmol) in DMF (3 mL). The reaction mixture isconcentrated, treated with a mixture of (1:1) DCM-TFA and stirred atroom temperature for 6 hours. The mixture is diluted with DCM and washedwith a solution of sat. NaHCO₃. The organic layers are combined, driedover MgSO₄, filtered and concentrated to afford a solid, which ispurified by silica gel column chromatography using 98:2 DCM:NH₃ (7M inMeOH). The residue is triturated with diethyl ether affording the titlecompound as a free base (70.5 mg, 59%). HPLC (254 nm): Rt 2.53 min(95%), m/z 433 (M+H)⁺.

Conversion into the mesylate salt, using2-[8-(4-morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-4,5-dihydro-thieno[2,3-c]pyrrol-6-one(69.2 mg, 016 mmol) and 0.1M methanesulfonic acid (1.598 mL), affordsthe title compound (76.2 mg, 90%). LCMS: Rt 2.50 min (96.4%), m/z (APCI)433 (M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO) δ (ppm) 2.34 (3H, s, MsOH),3.21-3.26 (4H, m), 3.83-3.86 (4H, m), 4.45 (2H, s), 7.17 (2H, d), 7.68(1H, br s), 7.74 (1H, s), 7.87-7.90 (3H, m), 8.30 (1H, s), 8.63 (1H, s),10.25 (1H, s).

Compound 199:4-{8-[4-(1-Isopropylpiperidin-4-yl)phenyl-amino]imidazo[1,2-a]pyrazin-5-yl}thiophene-2-carboxylicacid amide

This compound may be prepared using the same methods as described forCompound 202 using 2-(aminocarbonyl)thiophene-4-boronic acid in Step 4.

LCMS: Rt 0.88 min (100%) m/z (ESI) 461 (M+H)⁺.

Compound 200:2-Phenyl-N-{5-[5-(1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-ylamino]pyridin-2-yl}acetamide

Step 1: N-(5-Nitropyridin-2-yl)-2-phenylacetamide

A solution of 2-amino-5-nitropyridine (4.17 g, 30 mmol) in pyridine (30mL) is stirred at rt and a solution of phenylacetyl chloride (4.64 g, 30mmol) in THF (30 mL) is added dropwise. The mixture is stirred for 24 hand then poured into ice-water (250 mL) to afford a brown solid, whichis used without further purification.

Step 2:N-(5-Amino-pyridin-2-yl)-2-phenylacetamide

A solution of N-(5-nitropyridin-2-yl)-2-phenylacetamide (4.35 g, 16.9mmol) in EtOH (75 mL) and EtOAc (75 mL) is stirred at rt and ammoniumformate (4.27 g) and 10% Pd/C (500 mg) are added. The mixture is heatedat reflux for 30 min, cooled, filtered through celite and evaporated toafford the title compound as an off-white solid which was used withoutfurther purification.

Steps 3 and 4:2-Phenyl-N-{5-[5-(1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-ylamino]pyridin-2-yl}acetamide

This compound may be prepared using the same methods as described forcompound 184, steps 3 and 4. LCMS: Rt=0.99 min (100%), m/z (ESI) 411(M+H)⁺.

Compound 201:[6(4-Isopropylpiperazin-1-yl)pyridin-3-yl]-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]amine

This compound may be prepared using the same methods as described forCompound 184, using 6(4-isopropylpiperazin-1-yl)pyridin-3-ylamine inStep 3. LCMS: Rt=0.72 min (95%), m/z (ESI) 404 (M+H)⁺.

Compound 202:[4-(1-Isopropylpiperidin-4-yl)phenyl]-[5-(1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-yl]amine

Step 1: 1-Isopropyl-4-(4-nitrophenyl)piperidine

4-(4-Nitrophenyl)piperidine (250 mg, 1.21 mmol), K₂CO₃ (170 mg, 1.21mmol) and 2-iodopropane (240 μL, 2.4 mmol) are stirred in acetonitrile(3 mL) in a sealed tube at 120° C. for 45 min. The mixture is cooled andthe solvent removed under reduced pressure. The residue is partitionedbetween DCM (20 mL) and water (5 mL), the layers are separated and theDCM is washed with water (5 mL) and brine (5 mL) and dried over MgSO₄.Evaporation of the solvent affords the title compound (300 mg) which isused without further purification.

Step 2: 4-(1-Isopropylpiperidin-4-yl)phenylamine

Hydrazine (35% by weight in water, 0.67 mL, 7.2 nnol) and 10% Pd/C (38mg, 0.03 mmol) are added to a solution of1-isopropyl-4-(4-nitrophenyl)piperidine (180 mg, 0.72 mmol) in EtOH (10mL) and the mixture is heated at reflux for 3 h. After cooling, themixture is filtered through celite and the solvent evaporated. Theresidue is redissolved in DCM (25 mL), dried over MgSO₄ and the solventevaporated to afford the desired compound as a pale yellow solid (113mg, 0.52 mmol) which was used without further purification.

Step 3:(5-Bromo-imidazo[1,2-a]pyrazin-8-yl)-[4-(1-Isopropylpiperidin-4-yl)-phenyl]amine

N,N-Diisopropylethylamine (360 μL, 2.2 mmol) is added to a mixture of4-(1-isopropylpiperidin-4-yl)phenylamine (220 mg, 1.0 mmol) and5,8-dibromoimidazo[1,2-a]pyrazine (280 mg, 1.0 mmol) in ^(i)PrOH (5 mL)and heated at reflux for 48 h. The mixture is cooled and the solventevaporated under reduced pressure to afford an orange-brown solid. Thisis partitioned between DCM (50 mL) and water (20 mL) and the layers areseparated. The organic phase is washed with citric acid (10% aq., 3×25mL). The combined washings are extracted with DCM (25 mL) and then madebasic by addition of NaHCO₃ (s). The mixture is extracted with DCM (3×25mL) and the combined extracts dried over MgSO₄ and evaporated. Thematerial is used without further purification.

Step 4:[4-(1-Isopropylpiperidin-4-yl)phenyl]-[5-(1H-pyrazol-4-yl)imidazo[2-a]pyrazin-8-yl]amine

This step may be performed using the same methods as described forCompound 184, step 4, using (5-bromo-imidazo[1,2-a]pyrazin-8-yl)-[4-(1-isopropylpiperidin-4-yl)-phenyl]amine. LCMS: Rt 0.78 min (100%) m/z(ESI) 402 (M+H)⁺.

Compound 204:4-{8-[4-(4-Isopropyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-furan-2-carboxylicacid amide

In the same way as described for Compound 178, step 4, using,5-bromo-imidazo[1,2-a]pyrazin-8-yl-[4-(4-isopropyl-piperazin-1-yl)-phenyl]-amine(0.08 g, 0.19 mmol),4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-furan-2-carboxylic acidamide (0.092 g, 0.39 mmol), 1.5M Na₂CO₃ (1.02 mL, 1.53 mmol) andPd(PPh₃)₄ (0.055 g, 0.47 mmol) in dioxane (4 mL). Purification usingsilica gel column chromatography, eluting with DCM followed by 97:3DCM:NH₃ (7M in MeOH), affords the title compound (30.2 mg, 36%).

Conversion into the mesylate salt as described in for Compound 178, step4, using 1M methanesulfonic acid (0.068 mL), affords the title compoundas a yellow solid (31.6 mg, 86%). LCMS: Rt 1.84 min (90.3%); m/z (APCI)446 (M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO) δ (ppm) 1.35 (6H, d), 2.34 (3H, s,MsOH), 2.97-3.03 (2H, t), 3.22-3.36 (2H, m), 3.55-3.63 (3H, m), 3.86(2H, d) 7.06 (2H, d), 7.60 (1H, br s), 7.67 (2H, s), 7.79 (1H, s), 7.98(3H, br d), 8.17 (1H, s), 8.51 (1H, s), 9.28 (1H, br s), 9.60 (1H, s).

Compound 205:5-{8-[4-(4-Isopropyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-2,3-dihydro-isoindol-1-one

Step 1: 1-Isopropyl-4-(4-nitro-phenyl)-piperazine

To a solution of 4-fluoronitrobenzene (5 g, 35.4 mmol) in THF (50 mL),1-isopropylpiperazine (4.54 g, 35.4 mmol) and K₂CO₃ (7.35 g, 53.2 mmol)are added. The reaction mixture is stirred at room temperatureovernight. The solvent is removed in vacuo and the residue ispartitioned between EtOAc and water. The organic layer is washed withbrine, dried over MgSO₄, filtered and concentrated. The crude compoundis purified by silica gel column chromatography using 99:1 and 98:2DCM:NH₃ (7M in MeOH) to give the title compound (8.2 g, 94%).

Step 2: 4-(4-Isopropyl-piperazin-1-yl)-phenylamine

1-Isopropyl-4-(4-nitro-phenyl)-piperazine (8.3 g, 33.2 mmol) isdissolved in MeOH (120 mL) and tin (II) dichloride dihydrate (37.4 g,0.165 mol) is added. The mixture is cooled using a water bath and conc.HCl is added (36 mL). The reaction is stirred at room temperatureovernight. After removing the methanol, the resultant solution isbasified using conc. NaOH (pH 11). The water phase is extracted withdiethyl ether (3×) and the organic layers combined, dried over MgSO₄,filtered, concentrated in vacuo to afford the title compound (6.4 g,88%).

Step 3:(5-Bromo-imidazo[1,2-a]pyrazin-8-yl)-[4-(4-isopropyl-piperazin-1-yl)-phenyl]-amine

A solution of 5,8-dibromo-imidazo[1,2-a]pyrazine (0.134 g, 0.48 mmol),4-(4-isopropyl-piperazin-1-yl)-phenylamine (0.118 g, 0.54 mmol) anddiisopropylethylamine (0.093 mL, 0.54 mmol) is stirred at 90° C. for 18hours. The reaction mixture is concentrated in vacuo and partitionedbetween DCM and sat. NaHCO₃. The organic layer is washed with brine,dried over MgSO₄, filtered and concentrated. The residue is purified bysilica gel column chromatography eluting with 98:2 DCM:MeOH to affordthe title compound (0.086 g,. 43%).

Step 4:5-{8-[4-(4-Isopropyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-2,3-dihydro-isoindol-1-one

In the same way as described for Compound 178, step 4, using,(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-[4-(4-isopropyl-piperazin-1-yl)-phenyl]-amine(0.08 g, 0.19 mmol),5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-2,3-dihydro-isoindol-1-one(0.075 g, 0.29 mmol), 1.5M Na₂CO₃ (1.03 mL), and Pd(PPh₃)₄ (0.055 g,0.047 mmol) in dioxane. The reaction mixture is purified by silica gelcolumn chromatography eluting with DCM followed by 96:4 DCM:NH₃ (7M inMeOH). The residue is triturated using diethyl ether to afford the titlecompound (49.2 mg, 55%). Conversion into the mesylate salt, as describedin step 4, Example 178, using 1M methanesulfonic acid (0.1 mL), givesthe title compound (40.8 mg, 69%). LCMS: Rt 1.93 min (94.2%); m/z (APCI)468 (M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO) δ (ppm) 1.34 (6H, d), 2.34 (3H, s,MsOH), 3.04 (2H, t), 3.22 (2H, m), 3.57-3.63 (3H, m), 3.91 (2H, d), 4.52(2H, s), 7.13 (2H, d), 7.49 (1H, s), 7.81-7.94 (6H, m), 8.11 (1H, s),8.76 (1H, br s), 9.32 (1H, br s).

Compound 206:5-{8-[4-(1-Isopropylpiperidin-4-yl)phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-2,3-dihydroisoindol-1-one

This compound may be prepared using the same methods as described forcompound 202, using5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-2,3-dihydroisoindol-1-onein Step 4.

LCMS: Rt 0.86 min (100%) m/z (ESI) 467 (M+H)⁺.

Compound 208:N-(5-Morpholinopyridin-2-yl)-5-(1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-amine

Step 1: 4-(6-Nitro-pyridin-3-yl)-morpholine

A mixture of 5-bromo-2-nitro-pyridine (5 g, 24.75 mmol), tetrabutylammonium iodide (0.46 g, 1.24 mmol), K₂CO₃ (3.76 g, 27.22 mmol) andmorpholine (2.34 g, 27.22 mmol) in DMSO (50 mL) is stirred at 80° C. ina stem tube for 20 hours. The reaction mixture is diluted with ethylacetate and filtered. The organic solution is washed with water, driedover MgSO₄ and concentrated in vacuo. The residue is triturated withdichloromethane and hexane to afford the title compound as a solid (2.39g, 46%).

Step 2: 5-Morpholin-4-yl-pyridin-2-ylamine

In the same way as described in for Compound 154, step 2 using4-(6-nitro-pyridin-3-yl)-morpholine (2.39 g, 11.43 mmol), tin(II)chloride dihydrate (12.9 g, 57.17 mmol) in MeOH (40 mL) and conc. HCl(12.4 mL). The reaction mixture is concentrated in vacuo cooled to 0° C.and dissolved in water. The aqueous solution is basified using conc.NaOH and extracted with ethyl acetate (3×). The organic layers arecombined, dried over MgSO₄, filtered and concentrated to afford thetitle compound (1.77 g, 87%) which is used in the next step withoutfurther purification.

Step 3:(5-Bromo-imidazo[1,2-a]pyrazin-8-yl)-(5-morpholin-4-yl-pyridin-2-yl)-amine

In the same way as described for compound 90, step 1, using5,8-dibromo-imidazo[1,2-a]pyrazine (0.4 g, 1.45 mmol),5-morpholin-4-yl-pyridin-2-ylamine (0.258 g, 1.23 mmol), NaO^(t)Bu (0.19g, 2.04 mmol), Pd₂dba₃ (0.053 g, 0.058 mmol) and Xantphos (0.067 g,0.116 mmol) in toluene (6 mL). After removing the solvent in vacuo, themixture is purified by silica gel column chromatography eluting with 1:4petroleum ether:ethyl acetate followed by 99:1 DCM:NH₃ (7M in MeOH). Thetitle compound is isolated (216.4 mg, 47%). LCMS: Rt 2.16 min (93.2%).

Step 4:N-(5-Morpholinopyridin-2-yl)-5-(1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-amine

In the same way as described for compound 127, step 4 using(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-(5-morpholin-4-yl-pyridin-2-yl)-amine(100 mg, 0.266 mmol),4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole (77.6 mg,0.4 mmol), Pd(PPh₃)₄ (77 mg, 0.066 mmol) and 1.5M Na₂CO₃ (1.42 mL, 2.13mmol) in dioxane (4 mL). The crude compound is purified by silica gelcolumn chromatography eluting with 96:4 DCM:NH₃ (7M in MeOH) to affordthe title compound which is triturated with diethyl ether (12.9 mg, 8%)and converted into the mesylate salt (6.1 mg, 38%) LCMS: Rt 1.95 min(96%), m/z (APCI) 363 (M+H)⁺, ¹H-NMR (400 MHz, d₆-DMSO) δ (ppm) 2.34(3H, s, MsOH), 3.20 (4H, m), 3.83 (4H, m), 7.38 (1H, br s), 7.64 (1H, brs), 7.71 (1H, s), 7.86 (1H, d), 7.98 (1H, d), 8.04 (2H, m), 8.35 (2H, brs), 8.43 (1H, s).

Compound 209:4-{8-[6-(4-Isopropyl-piperazin-1-yl)-pyridin-3-ylamino]-imidazo[1,2-a]pyrazin-5-yl}-furan-2-carboxylicacid amide

Step 1: 4-Bromo-furan-2-carboxylic acid amide

To a solution of 4,5-dibromo-furan-2-carboxylic acid (7.79 g, 28.85mmol) in NH₄OH (100 ml) is added zinc dust (2.29 g, 34.62 mmol) in smallportions. The reaction mixture is stirred at room temperature for 7minutes then filtered over celite and washed with water and 2M HCl. Thefiltrate is acidified to pH I using collc. HCl and extracted with ethylacetate (3×). The organic phase is washed with brine, dried over MgSO₄,filtrated and concentrated in vacuo to give an oil (4.96 g) whichsolidifies on standing to give a white solid, used in the amideformation step, without further purification.

The solid (4.93 g, 25.81 mmol) is dissolved in thionyl chloride (44.2mL) and refluxed for 1 hour. After removing the solvent in vacuo theresidue is dissolved in dichloromethane (75 mL) and a solution of 0.5 MNH3 in dioxane (52 mL) is added. The reaction mixture is stirred at roomtemperature for 1 hour, then 33% aq. NH₃ (5 mL) is added and thereaction stirred for additional 2 hours. The solvent is removed in vacuoand the residue taken-up with a solution of sat. NaHCO₃. The basicsolution is extracted using ethyl acetate (3×), the combined organiclayers are dried over MgSO₄ and concentrated in vacuo. Purification bysilica gel column chromatography eluting with a mixture of (50:49:1)ethyl acetate:petroleum ether:acetic acid, affords the title compound(1.2 g, 22%).

Step 2:4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-furan-2-carboxylic acidamide

4-Bromo-furan-2-carboxylic acid amide (1.2 g, 6.32 mmol),bis(pinacolato)diboron (1.76 g, 6.94 mmol), PdCl₂dppf (0.154 g, 189 mol)and KOAc (1.85 g, 18.94 mmol) are suspended in dioxane (20 mL), purgedwith nitrogen for 5 minutes and then heated at 90° C. overnight. Thesolvent is removed in vacuo and the residue partitioned between ethylacetate and water. The aqueous layer is extracted three times with ethylacetate and the combined organic phases are washed with brine, filteredthrough MgSO₄ and evaporated. The solid residue is triturated withhexane and dried in vacuo to afford the title compound as a solid (0.984g, 66%).

Step 3:4-{8-[6-(4-Isopropyl-piperazin-1-yl-pyridin-3-ylamino]-imidazo[1,2-a]pyrazin-5-yl}-furan-2-carboxylicacid amide

In the same way as described for Compound 178, step 4, using(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-[6-(4-isopropyl-piperazin-1-yl)-pyridin-3-yl]-amine(0.05 g, 0.12 mmol),4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-furan-2-carboxylic acidamide (0.050 g, 0.22 mmol), 1.5M Na₂CO₃ (0.64 mL, 0.96 mmol) andPd(PPh₃)₄ (0.042 g, 0.36 mmol) in dioxane (2 mL). Purification usingsilica gel column chromatography, eluting with DCM followed by 99:1 and97:3 DCM:NH₃ (7M in MeOH), affords the title compound (38 mg, 70%).

Conversion into the mesylate salt as described for Compound 178, step 4,affords the title compound as a yellow solid (28 mg, 52%). LCMS: Rt 2.58min (95%); m/z (ES⁺) 447 (M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO) δ (ppm) 1.34(6H, d), 2.36 (6H, s, 2×MsOH), 3.06-3.20 (4H, m), 3.52-3.59 (3H, m),4.42-4.46 (2H, d), 7.12 (1H, d), 7.63 (1H, br s), 7.68 (2H, d), 7.84(1H, s), 8.01 (1H, br sH), 8.21 (1H, s), 8.27 (1H, m), 8.53 (1H, s),8.82 (1H, br s), 9.37 (1H, br s), 9.94 (1H, br s).

Compound 210:5-{8-[6-(4-Isopropyl-piperazin-1-yl)-pyridin-3-ylamino]-imidazo[1,2-a]pyrazin-5-yl}-2,3-dihydro-isoindol-1-one

Step 1: 1-Isopropyl-4-(5-nitro-pyridin-2-yl)-piperazine

To a solution of 2-chloro-5-nitropyridine (2.5 g, 15.7 mmol) in THF (25mL), are added 1-isopropylpiperazine (2.01 g, 15.7 mmol) and K₂CO₃ (3.25g, 23.6 mmol). The reaction mixture is stirred 50° C. for 4 hours andthen at 70° C. overnight. The solvent is removed in vacuo and theresultant orange solid is triturated using 10:1 petroleum ether-diethylether. The isolated compound (3.7 g, 94%) is used in the next stepwithout further purification.

Step 2: 6-(4-Isopropyl-piperazin-1-yl)-pyridin-3-yl-amine

1-Isopropyl-4-(5-nitro-pyridin-2-yl)-piperazine (0.9 g, 3.6 mmol) isdissolved in MeOH (20 mL) and tin (II) dichloride dihydrate (4 g, 18mmol) is added. The mixture is cooled using a water bath and conc. HClis added (4 mL). The reaction is stirred at room temperature overnight.After removing the methanol, the resultant light yellow solution isbasified using conc. NaOH (pH 11) and a white precipitate is formed. Thesolid is collected by filtration and the water is extracted with diethylether (5×). The organic layers are combined, dried over MgSO₄, filtered,concentrated under vacuum to afford an orange oil which crystallizes onstanding to afford an orange solid (0.68 g, 86%).

Step 3:(5-Bromo-imidazo[1,2-a]pyrazin-8-yl)-[6-(4-isopropyl-piperazin-1-yl)-pyridin-3-yl]-amine

To a solution of 5,8-dibromo-imidazo[1,2-a]pyrazine (0.85 g, 3.1 mmol)in isopropanol are added6-(4-isopropyl-piperazin-1-yl)-pyridin-3-yl-amine (0.68 g, 3.1 mmol) anddiisopropylethylamine (0.6 mL, 4.6 mmol) and the mixture is stirred at90° C. for 48 hours. The solvent is removed in vacuo and the producttaken up in DCM-10% citric acid. The organic layer is discarded and theaqueous solution basified with sat. NaHCO₃ (pH 7-8) and extracted usingDCM. The organic layer is dried over MgSO₄, filtered and concentrated invacuo to afford the title compound as a purple solid (0.92 g, 71%). HPLC(254 nm): Rt 3.30 min (95.8%); m/z 416/418 (M+H)⁺.

Step 4:5-{8-[6-(4-Isopropyl-piperazin-1-yl)-pyridin-3-ylamino]-imidazo[1,2-a]pyrazin-5-yl}-2,3-dihydro-isoindol-1-one

In the same way as described for Compound 178, step 4, using(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-[6-(4-isopropyl-piperazin-1-yl)-pyridin-3-yl]-amine(0.05 g, 0.12 mmol),5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-2,3-dihydro-isoindol-1-one(0.056 g, 0.22 mmol), 1.5M Na₂CO₃ (0.64 mL, 0.96 mmol) and Pd(PPh₃)₄(0.042 g, 0.36 mmol) in dioxane (2 mL). Purification using silica gelcolumn chromatography, eluting with DCM followed by 99:1 and 97:3DCM:NH₃ (7M in MeOH), affords the title compound as a yellow solid (28mg, 50%). Conversion into the mesylate salt, as described for Compound178, step 4, affords the title compound as a yellow solid (32 mg, 95%).LCMS: Rt 2.70 min (92%); m/z (APCI) 469 (M+H)⁺; ¹H-NMR (400 MHz,d₆-DMSO) δ (ppm) 1.34 (6H, d), 2.34 (3H, s, MsOH), 3.10-3.21 (4H, m),3.57-3.59 (3H, m), 4.43 (2H, d), 4.52 (2H, s), 7.08 (1H, d), 7.52 (1H,s), 7.77-7.94 (4H, m), 8.07 (1H, s), 8.30 (1H, d), 8.75 (1H, s), 8.83(1H, br s), 9.34 (1H, br s), 9.88 (1H, br s).

Compound 212:4-[8-(4-Morpholin-4-yl-phenylamino)-imidazol[1,2-a]pyrazin-5-yl]-2H-pyrazole-3-carbonitrile

Step 1: 4-Bromo-2-(tetrahydro-pyran-2-yl)-2H-pyrazole-3-carbonitrile

4-Bromo-1H-pyrazole-3-carbonitrile (150 mg, 0.87 mmol) is dissolved in3,4-dihydro-2H-pyran (25 mL, 2.6 mmol) in the presence of a catalyticamount of TFA (1 mg, 0.009 mmol). The reaction mixture is stirred at 95°C. for 1 hour, cooled and then quenched using NaH (1.2 mg, 0.052 mmol).After removing the solvent, the residue is purified by silica gel columnchromatography eluting with a mixture of 3:1 petroleumether-dichloromethane, followed by 10:1 petroleum ether-ethyl acetate.The fractions containing the desired compounds are collected andconcentrated in vacuo to afford the title compound as a colorless oil(90 mg, 40%).

Step 2:(4-Morpholin-4-yl-phenyl)-(5-tributylstannanyl-imidazo[1,2-a]pyrazin-8-yl)-carbamicacid tert-butyl ester

To a cooled (−78° C.) solution of(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-(4-morpholin-4-yl-phenyl)-carbamicacid tert-butyl ester (2.55 g, 5.38 mmol) in tetrahydrofuran (20 mL) isadded isopropylmagnesium chloride (0.829 g, 8.06 mmol). After stirringfor 5 min tributyltin chloride (2.97 g, 9.14 mmol) is added and thereaction is stirred at −78 ° C. for 10 min before being allowed to warmto room temperature. After stirring at room temperature for 30 min, themixture is concentrated. The residue is purified by silica gel flashcolumn chromatography eluting with 5:1 petroleum ether-ethyl acetatefollowed by 1:1 petroleum ether-ethyl acetate to afford the product asyellow solid (2.35 g, 64%).

Step 3:4-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-2H-pyrazole-3-carbonitrile

(4-Morpholin-4-yl-phenyl)-(5-tributylstannanyl-imidazo[1,2-a]pyrazin-8-yl)-carbamicacid tert-butyl ester (385 mg, 0.56 mmol),4-bromo-2-(tetrahydro-pyran-2-yl)-2H-pyrazole-3-carbonitrile (90 mg,0.35 mmol) and Pd(PPh₃)₄ (41 mg, 0.035 mmol) are suspended in DMF and,after purging with nitrogen, the reaction mixture is stirred at 90° C.overnight. The solvent is evaporated in vacuo and the resulting yellowoil (590 mg) is dissolved in a mixture 1:1 TFA-DCM (2 mL) (2 drops H₂O)and stirred at room temperature overnight. The mixture is diluted withDCM and washed with a solution of sat. NaHCO₃. The organic layer isdried over MgSO₄, filtered and concentrated to afford a yellow solid.Purification by silica gel column chromatography using DCM followed by98:2 and 96:4 DCM:NH₃ (7M in MeOH) affords an oil, which is purified byreverse phase preparative HPLC to give the title compound (5 mg, 2%).LCMS: Rt 2.81 min (92%); m/z (ES⁺) 387 (M+H)⁺; ¹H-NMR (400 MHz, d₆-DMSO)δ (ppm) 3.10 (4H, m), 3.79 (4H, m), 6.98 (2H, d), 7.52 (1H, s), 7.73(1H, s), 7.93 (3H, d), 7.99 (1H, s), 8.55 (1H, s), 9.58 (1H, s).

Compound 217:2-[8-(4-Morpholin-4-yl-phenylamino)-imidazol[1,2-a]pyrazin5-yl]-5,6-dihydro-furo[2,3-c]pyrrol-4-one

Step 1: 5-Bromo-2-bromomethyl-furan-3-carboxylic acid methyl ester

To a cooled solution (0° C.) of 2-methyl-furan-3-carboxylic acid methylester (3 g, 21.43 mmol) in DMF (60 mL) is added NBS (8.77 g, 49.28 mmol)in portions. The reaction mixture is stirred at 0° C. for 1 hour and atroom temperature overnight. The reaction mixture is then diluted withwater and extracted with diethyl ether (3×). The organic layers arecombined and washed with brine and water, dried over MgSO₄, filtered andconcentrated in vacuo. The crude compound is purified by silica gelcolumn chromatography eluting with 95:5 petroleum ether:diethyl ether toafford the title compound (0.626 g, 10%).

Step 2: 5-Bromo-2-[(4-methoxy-benzylamino)-methyl]-furan-3-carboxylicacid methyl ester

A solution of 5-bromo-2-bromomethyl-furan-3-carboxylic acid methyl ester(1.74, 5.88 mmol) and p-methoxy benzylamine (0.76 mL, 5.88 mmol) in MeOH(30 mL) is stirred at 85° C. for 1.5 hours. The reaction mixture isconcentrated in vacuo and the residue purified by silica gel columnchromatography eluting with 4:1 petroleum ether:ethyl acetate followedby 1:1 petroleum ether:ethylacetate. The title compound is isolated(643.8 mg).

Step 3: 5-Bromo-2-[(4-methoxy-benzylamino)-methyl]-furan-3-carboxylicacid

A solution of5-bromo-2-[(4-methoxy-benzylamino)-methyl]-furan-3-carboxylic acidmethyl ester (643.8 mg, 1.82 mmol) and LiOH (115 mg, 2.74 mmol) in 1:1THF:H₂O (30 mL) is refluxed for 3 hours. The solvent is evaporated invacuo to afford the title compound (620 mg, 99%) which is used in thenext step without further purification.

Step 4: 2-Bromo-5-(4-methoxy-benzyl)-3a, 5,6, 6a-tetrahydro-furo[2,3-c]pyrrol-4-one

A solution of5-bromo-2-[(4-methoxy-benzylamino)-methyl]-furan-3-carboxylic acid (618mg, 1.82 mmol) and SOCl₂ (0.136 mL) in DCM (2 mL) is stirred at 85° C.for 18 hours. The mixture is concentrated in vacuo and purified bysilica gel column chromatography eluting with 7:3 petroleum ether:ethylacetate to afford the title compound (0.169 g, 28%).

Step 5: 2-Bromo-3a, 5,6,6a-tetrahydro-furo[2,3-c]pyrrol-4-one

A solution of2-bromo-5-(4-methoxy-benzyl)-3a,5,6,6a-tetrahydro-furo[2,3-c]pyrrol-4-one(0.16 g, 0.498 mmol) in TFA (1.8 mL) and anisole (0.2 mL) is stirred at80° C. for 2.5 hours. The reaction mixture is then diluted with ethylacetate and neutralised with sat. NaHCO₃. The aqueous layer is extractedwith ethyl acetate (3×) and the combined organic layers are dried overMgSO₄, filtered and concentrated in vacuo. The crude compound ispurified by silica gel column chromatography eluting with 1:1 petroleumether:ethyl acetate followed by 1:4 petroleum ether:ethyl acetate, toafford the title compound (79 mg, 78%).

Step 6:2-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin5-yl]-5,6-dihydro-furo[2,3-c]pyrrol-4one

In the same way as described for Compound 212, step 3 using(4-morpholin-4-yl-phenyl)-(5-tributylstannanyl-imidazo[1,2-a]pyrazin-8-yl)-carbamicacid tert-butyl ester (255 mg, 0.373 mmol),2-bromo-3a,5,6,6a-tetrahydro-furo[2,3-c]pyrrol-4-one (50 mg, 0.249 mmol)and Pd(PPh₃)₄ (29 mg, 0.0249 mmol) in DMF (3 mL). The reaction mixtureis concentrated in vacuo and the residue dissolved in 1:1 DCM:TFA (dropof H₂O) and stirred at room temperature for 3 hours. The mixture ispartitioned between ethyl acetate and sat. NaHCO₃ and the aqueous layeris extracted with ethyl acetate (3×). The organic layers are combined,dried over MgSO₄, filtered and concentrated in vacuo. The residue ispurified by silica gel column chromatography eluting with DCM followedby a 97:3 mixture DCM:NH₃ (7M in MeOH). The fractions containing thedesired compound are concentrated in vacuo to give a solid which istriturated with diethyl ether to afford the title compound (66.3 mg,64%). Conversion of the compound into the mesylate salt using 1M MsOH(0.16 mL) affords the title product as a solid (69.3 mg, 87%). LCMS: Rt2.52 min (99%), m/z (APCI) 417 (M+H)⁺, ¹H-NMR (400 MHz, d₆-DMSO) δ(ppm). 2.36 (3H, s, MsOH), 3.25 (4H, m), 3.84 (4H, m), 4.52 (2H, s),7.18 (2H, br s), 7.40 (1H, s), 7.83 (1H, s), 7.90 (3H, m), 8.21 (1H, brs), 8.44 (1H, s), 10.25 (1H, br s).

Compound 218:(3-Dimethylaminomethyl-4-morpholin-4-yl-phenyl)-[5-(1H-pyrazol-4-yl)-imidazol[1,2-a]pyrazin-8-yl]-amine

Step 1: (2-Morpholin-4-yl-5-nitro-phenyl)-methanol

To a cooled (0° C.) solution of 2-morpholin-4-yl-5-nitro-benzaldehyde(0.8 g, 3.39 mmol) in MeOH (5 mL) is added NaBH₄ (0.125 g, 3.39 mmol)and the reaction mixture is stirred at room temperature for 3 hours.After quenching the reaction with water, the solvent is removed in vacuoand the residue dissolved in ethyl acetate and washed with brine. Theorganic layer is dried over MgSO₄, filtered and concentrated to affordthe title compound (870 mg), which is used in the next step withoutfurther purification.

Step 2: (5-Amino-2-morpholin-4-yl-phenyl)-methanol

To a solution of (2-morpholin-4-yl-5-nitro-phenyl)-methanol (870 mg) inethanol (40 mL), palladium hydroxide (87 mg) is added and the mixture isstirred in a Parr-apparatus under hydrogen pressure (10 bars) for 4hours. The reaction mixture is filtrated over Celite 521, washed withethanol and concentrated in vacuo to give the title compound (640 mg,83%).

Step 3:3-(tert-Butyl-dimethyl-silanyloxymethyl)-4-morpholin-4-yl-phenylamine

A solution of (5-amino-2-morpholin-4-yl-phenyl)-methanol (640 mg, 3.07mmol), tert-butyldimethylsilyl chloride (509 mg, 3.38 mmol) andimidazole (250 mg, 3.68 mmol) in dimethylformamide (20 mL) is stirred atroom temperature overnight. The solvent is removed in vacuo and theresidue partitioned between water and ethyl acetate. The organic layeris washed with brine, dried over MgSO₄, filtered and concentrated toafford a crude product. Purification, using silica gel columnchromatography, eluting with DCM followed by a 95:5 mixture DCM:MeOH,affords the title compound as a pink solid (390 mg, 27%).

Step 4:(5-Bromo-imidazo[1,2-a]pyrazin-8-yl)-[3-(tert-butyl-dimethyl-silanyloxymethyl)-4-morpholin-4-yl-phenyl]-amine

In the same way as described in the general procedure for aminedisplacement using 5,8-dibromo-imidazo[1,2-a]pyrazine (0.534 g, 1.94mmol),3-(tert-butyl-dimethyl-silanyloxymethyl)-4-morpholin-4-yl-phenylamine(0.751 g, 2.33 mmol), N,N-diisopropyethylamine (0.31 mL, 1.8 mmol) andiso-propanol (10 mL). Purification by silica gel column chromatographyeluting with 7:3 petroleum ether:ethyl acetate followed by triturationwith diethyl ether affords the title compound (0.216 g, 17%).

Step 5.(5-Bromo-imidazo[1,2-a]pyrazin-8-yl)-[3-(tert-butyl-dimethyl-silanyloxymethyl)-4-morpholin-4-yl-phenyl]-carbamicacid tert-butyl ester

In the same way as described in the general procedure for Boc protectionusing (5-bromo-imidazo[1,2-a]pyrazin-8-yl)-[3-(tert-butyl-dimethyl-silanyloxymethyl)-4-morpholin-4-yl-phenyl]-amine(0.216 g, 0.41 mmol), di-tert-butyl-dicarbonate (0.266 g, 1.21 mmol) andN,N-dimethylaminopyridine (4 mg, 0.04 mmol) in DCM (5 mL). The titlecompound (0.164 g, 54%) is isolated after purification by silica gelcolumn chromatography eluting with 4:1 petroleum ether:ethyl acetatefollowed by 7:3 petroleum ether:ethyl acetate.

Step 6:(5-Bromo-imidazo[1,2-a]pyrazin-8-yl)-(3-hydroxymethyl-4-morpholin-4-yl-phenyl)-carbamicacid tert-butyl ester

A solution of(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-[3-(tert-butyl-dimethyl-silanyloxymethyl)-4-morpholin-4-yl-phenyl]-carbamicacid tert-butyl ester (0.164 g, 0.265 mmol) in 1M tetrabutylammoniumfluoride in THF (0.29 mL, 0.29 mmol) is stirred at room temperatureovernight. The solvent is removed under vacuum and the residue ispurified by silica gel column chromatography eluting with ethyl acetate.The title compound is isolated (0.126 g, 94%).

Step 7: Methanesulfonic acid5-[(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-tert-butoxycarbonyl-amino]-2-morpholin-4-yl-benzylester

To a cooled solution (0° C.) of(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-(3-hydroxymethyl-4-morpholin-4-yl-phenyl)-carbamicacid tert-butyl ester (0.17 g, 0.34 mmol) in DCM (0.5 mL), is addedtriethylamine (0.104 mL, 0.74 mmol) followed by methanesulfonyl chloride(0.046 mL, 0.6 mmol). The reaction mixture is stirred at 0° C. for 1hour and an additional hour at room temperature. The reaction mixture isdiluted with ethyl acetate and washed with water. The organic layer isdried over MgSO₄, filtered and concentrated in vacuo to give the titlecompound (0.224 g) used in the next step without further purification.

Step 8:(5-Bromo-imidazo[1,2-a]pyrazin-8-yl)-(3-dimethylaminomethyl-4-morpholin-4-yl-phenyl)-carbamicacid tert-butyl ester

To a solution of methanesulfonic acid5-[(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-tert-butoxycarbonyl-amino]-2-morpholin-4-yl-benzylester (0.098 g, 0.169 mmol) in THF (0.5 mL) is added 2MN,N-dimethylamine solution in THF (0.50 mL, 1.1 mmol) followed by K₂CO₃(0.028 g, 0.202 mmol). The reaction mixture is stirred at roomtemperature for 1.5 hours and at 75° C. for an additional 3 hours. Thesolvent is removed under vacuum and the residue is partitioned betweenethyl acetate and water. The organic layer is dried over MgSO₄, filteredand concentrated in vacuo to give the crude compound which is purifiedby silica gel column chromatography. Elution using 97:3 DCM:NH₃ (7M inMeOH) affords the title compound (0.0543 g, 61%). LCMS: Rt 2.37 min(95%), m/z (ES⁺) 531/533 (M+H)⁺.

Step 9:(3-Dimethylaminomethyl-4-morpholin-4-yl-phenyl)-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine

In the same way as described for Compound 85, step 1, using(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-(3-dimethylaminomethyl-4-morpholin-4-yl-phenyl)-carbamicacid tert-butyl ester (54 mg, 0.1024 mmol),4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole (29.8 mg,0.153 mmol), Pd(PPh₃)₄ (12 mg, 0.0102 mmol) and NaO^(t)Bu (39 mg, 0.41mmol) in 3:1 DMF/water (4 mL). The reaction mixture is concentratedunder reduced pressure and the residue is dissolved in a mixture 1:1DCM:TFA (drop of water). After stirring the solution at room temperaturefor 4 hours, ethyl acetate is added and the mixture is basified usingsat. NaHCO₃. The organic layer is separated, dried over MgSO₄, filteredand concentrated in vacuo to afford a crude compound. Purification bysilica gel column chromatography eluting with DCM followed by 95:5DCM:NH₃ (7M in MeOH), affords the title compound (28.1 mg, 66%).Conversion into the mesylate salt using 0.1M MsOH (0.67 mL) yields thetitle compound (26.9 mg, 78%). LCMS: Rt 1.97 min (97%), m/z (APCI) 419(M+H)⁺, ¹H-NMR (400 MHz, d₆-DMSO) δ (ppm) 2.35 (3H, s, MsOH), 2.74-2.89(10H, m), 3.82 (4H, m), 4.39 (2H, d), 7.42 (1H, d), 7.61 (1H, s), 7.81(1H, s), 8.08 (1H, d), 8.14-8.26 (4H, m), 9.19 (1H, br s), 9.75 (1H, brs).

Compound 220:7-Fluoro-5-{8-[4-(4-isopropyl-piperazin-1-yl)phenylamino]imidazo[1,2-a]pyrazin-5-yl}-2,3-dihydroisoindol-1-one

Step 1: 4-Bromo-2,6-difluorobenzoic acid methyl ester

To a suspension of 4-Bromo-2,6-difluoro-benzoic acid (5 g, 21 mmol) inDCM (10 mL) is added thionyl chloride (15 mL) and DMF (0.5 mL). Themixture is stirred at room temperature for 2.5 h. It is then cooled to0° C. and MeOH (20 mL) is added carefully causing vigorous HClevolution. After stirring for an additional 0.5 h, the clear solution ispartitioned between DCM (50 mL) and water (50 mL). The organic layer iswashed with saturated NaHCO₃, brine, dried over MgSO₄ and the solvent isremoved under vacuum to afford the title compound as a pale yellow oilwhich is used without further purification.

Step 2: 4-Bromo-2-fluoro-6-(nitromethyl)benzoic acid methyl ester

Nitromethane (10 mL, 169 mmol, 8 equiv.) is added cautiously to asuspension of sodium hydride (4.05 g, 169 mmol, 8 equiv.) and MgSO₄ (40g) in DMSO (100 mL) at rt and the resulting slurry is stirred for 0.25h. To the resulting yellow slurry is added 4-bromo-2,6-difluorobenzoicacid methyl ester (5.3 g, 21 mmol) and the mixture is stirred at rt for3 days at which point all the starting material has been consumed. Water(200 mL) and 6M HCl (50 mL) is added followed by DCM (200 mL). Morewater (500 mL) is added to yield a clear biphasic system. The aqueouslayer is extracted with DCM (3×100 mL), the DCM layers are thencombined, washed with saturated NaHCO₃ and brine, and dried over MgSO₄.Evaporation of the solvent affords an orange solid containing 64% of thedesired material, used without further purification. LCMS; Rt=1.27 min(64%).

Step 3: 5-Bromo-7-fluoro-2,3-dihydro-isoindol-1-one

Crude 4-bromo-2-fluoro-6-(nitromethyl)benzoic acid methyl ester from theprevious step is dissolved in MeOH (100 mL). To this clear orangesolution is added Zinc dust (3.35 g, 51.3 mmol, 3 equiv.) followed byammonium formate (3.23 g, 51.3 mmol, 3 equiv.) which results in anexothermic reaction. After 0.3 h, 7M NH₃ in MeOH (50 mL) is added andthe mixture is stirred overnight at room temperature. The resultingmixture is filtered through celite, and the yellow filtrate is adsorbedon silica and roughly cleaned by LC using 94/6 DCM/ MeOH 7M NH₃. Removalof the solvent affords a tan solid that is redissolved in DCM, washedwith 10% NaOH, and concentrated under reduced pressure to leave a whitesolid, which is triturated further with small amounts of DCM to affordthe title compound. LCMS: Rt 0.96 min (100%) m/z 230/232 (M+H)⁺.

Step 4:7-Fluoro-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-2,3-dihydroisoindol-1one

A solution of 5-bromo-7-fluoro-2,3-dihydroisoindol-1-one (531 mg, 2.31mmol) in dioxane (5 mL) is stirred under nitrogen. Pd(dppf)Cl₂ (94 mg, 5mol %), potassium acetate (453 mg, 4.62 mmol, 2.0 equiv.) andbispinacolatodiboron (1.17 g, 4.62 mmol, 2 equiv.) are added, and thereaction is heated at 80° C. for 3 h. The resulting orange suspension isdiluted with DCM, filtered through celite and concentrated under vacuumto afford an oil that is redissolved in a minimum of DCM. Diethyl etheris slowly added to afford the title compound as a tan solid. LCMS: Rt1.18 min (100%) m/z 277/279 (M+H)⁺.

Step 5:7-Fluoro-5-{8-[4-(4-isopropyl-piperazin-1-yl)phenylamino]imidazo[1,2-a]pyrazin-5-yl}-2,3-dihydroisoindol-1-one

This compound may be prepared using the methods as described forCompound 184, step 4, using(5-bromo-imidazo[1,2-a]pyrazin-8-yl)-[4-(4-isopropylpiperazin-1-yl)phenyl]amineand the above boronate. LCMS: Rt=0.82 min (95%), m/z 486 (M+H)⁺.

Compound 224:2-(4-{4-[5-(1H-Pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-ylamino]phenyl}piperidin-1-yl)acetamide

This compound may be prepared using the methods described for Compound202, using 2-bromoacetamide in Step 1. LCMS: Rt=0.73 min (100%), m/z(ESI) 417 (M+H)⁺.

Compound 225:2-(4-{4-[5-(3-Methyl-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-ylamino]phenyl}piperidin-1-yl)acetamide

This compound may be prepared using the methods as described forCompound 224, using3-methyl-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole inStep 4. LCMS: Rt=0.75 min (100%), m/z (ESI) 431 (M+H)⁺.

Compound 226:2-(4-{4-[5-(1-Oxo-2,3-dihydro-1H-isoindol-5-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-phenyl}piperidin-1-yl)acetamide

This compound may be prepared using the methods as described forCompound 224, using5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-2,3-dihydroisoindol-1-onein Step 4. LCMS: Rt=0.79 min (100%), m/z (ESI) 483 (M+H)⁺.

Compound 229:5-{8-[4-(4-Isopropylpiperazin-1-yl)-3-trifluoromethylphenylamino]imidazo[1,2-a]pyrazin-5-yl}-2,3-dihydroisoindol-1-one

This compound may be prepared using the methods as described forCompound 206, using4-(4-isopropylpiperazin-1-yl)-3-(trifluoromethyl)phenylamine in Step 3.LCMS: Rt=0.98 min (95%), m/z 536 (M+H)⁺.

Purifications Conditions and Characterization

Routinely, post-synthesis all compounds may be purified using reversephase HPLC using a Gilson preparative HPLC system (322 pump, 155 UV/VISdetector, 215 liquid handler). The Gilson 215 acts as both auto-samplerand fraction collector. Compounds can also be purified by flashchromatography on silica gel.

Compounds are characterised by mass spectrometry using single quadrupoleinstrumentation with an electrospray source.

Biological Assays Demonstrating the Usefulness of the Compounds

EXAMPLE 1 MAPKAP-K5 Assay

MAPKAP-K5 reactions are performed in FlashPlate format using 0.1 or 0.2μCi 33P-ATP; 0.6 μM ATP; 1mU MAPKAP-K5; 3 μM MAPKAP-K5 peptidesubstrate, incubated at room temperature for 30 minutes.

Flashplate Assay:

The MAPKAP-K5 kinase reaction is performed in a 384 well polypropyleneplate (Matrix Technologies) and then transferred to astreptavidin-coated 384 well flashplate (Perkin-Elmer).

To wells containing 2 μL test compound or standard inhibitor, 13 μLEnzyme mix or diluent are added using a Hydra (Robbins Scientific).

Reactions are started by addition of 10 μL of [2.5×]substrate cocktailusing a Multidrop (Thermo-Labsystems), to give final concentrations inthe assay of:

1 mU MAPKAP-K5

3 μM MAPKAP-K5 peptide substrate

˜0.6 μM ATP

0.004 μCi [33P]-γ-ATP/μL

1× reaction buffer

Plates are incubated at room temperature for 30 minutes.

Reactions are terminated by the addition of 25 μL EDTA (50 mM) to eachwell using a Micro-fill (Biotek).

Reactions are transferred to a streptavidin-coated flashplate using aZymark robotic system. Plates are incubated for 60 minutes at roomtemperature.

All wells are washed 3 times with 100 μL phosphate buffered saline usinga Tecan plate washer.

Radioactivity is determined by scintillation counting of the flashplate(empty wells) on a Packard TopCount.

Enzyme Mix:

Enzyme

50 mM Tris HCl (pH 7.5)

0.1 mM EGTA

2 mM DTT

1 mg/ml BSA

Reaction Buffer:

50 mM Tris HCl (pH 7.5)

0.1 mM EGTA

10 mM Magnesium acetate

2 mM DTT

Exemplary Compounds of the Invention

The following compounds have been or can be prepared according to thesynthetic methods described above. For the purpose of Table 1 below,activity of each compound, which can be determined using the MAPKAPK5assay method described in Example 1, is expressed as follows:

TABLE 1 Structure and activity of of Exemplary Compounds CompoundMAPKAPK5 ID STRUCTURE MW IC₅₀ (nM) 1

361.41 ++ 2

416.49 + 3

403.49 + 4

416.49 + 5

389.46 + 6

388.48 + 7

388.48 + 8

375.44 + 9

347.43 ++ 10

319.33 + 11

319.33 +++ 12

377.45 + 13

414.47 + 14

374.45 + 15

291.32 + 16

306.33 + 17

328.74 + 18

344.38 + 19

359.39 + 20

334.34 + 21

410.48 + 22

376.42 + 23

320.31 + 24

418.46 + 25

298.35 + 26

427.51 ++ 27

315.34 ++ 28

368.40 + 29

450.52 + 30

369.41 ++ 31

333.36 ++ 32

418.51 +++ 33

333.36 + 34

335.76 + 35

410.44 ++ 36

355.38 +++ 37

398.22 + 38

320.32 + 39

433.45 + 40

430.47 + 41

395.43 + 42

277.29 + 43

428.50 + 44

464.49 + 45

389.42 ++ 46

471.57 + 47

472.55 + 48

448.53 + 49

416.44 + 50

418.51 + 51

320.31 + 52

373.37 + 53

432.49 +++ 54

486.53 + 55

388.44 +++ 56

390.45 +++ 57

402.46 +++ 58

432.53 + 59

432.53 + 60

496.58 + 61

363.38 ++ 62

363.38 + 63

339.36 + 64

402.46 +++ 65

474.57 + 66

338.37 + 67

362.40 + 68

420.48 + 69

454.56 +++ 70

438.51 + 71

354.39 +++ 72

429.53 + 73

434.57 + 74

418.50 + 75

512.54 + 76

486.58 + 77

479.59 + 78

434.57 + 79

506.63 + 80

462.52 + 81

446.56 + 82

436.50 ++ 83

436.50 +++ 84

419.49 ++ 85

388.43 +++ 86

429.53 + 87

430.52 + 88

432.53 + 89

452.52 + 90

425.45 ++ 91

362.40 + 92

333.36 + 93

424.47 +++ 94

462.52 ++ 95

438.50 + 96

330.35 + 97

407.45 + 98

507.62 + 99

445.53 +++ 100

376.42 + 101

377.45 + 102

401.47 +++ 103

419.49 ++ 104

463.50 + 105

419.49 + 106

433.52 + 107

416.49 + 108

437.46 ++ 109

401.47 + 111

415.50 + 112

452.48 +++ 114

494.56 +++ 115

481.61 + 116

441.54 + 117

467.58 + 118

433.54 +++ 119

388.48 +++ 120

441.54 + 121

428.50 + 122

445.50 ++ 123

445.50 ++ 124

461.96 + 125

403.47 + 126

470.58 + 127

433.54 +++ 128

428.50 + 129

428.50 + 130

428.50 + 131

375.44 + 132

400.49 + 133

463.43 + 134

396.41 + 135

402.46 + 136

461.96 +++ 137

455.57 +++ 138

391.50 + 139

417.47 + 140

463.49 +++ 141

410.44 + 142

455.57 + 143

428.50 + 144

412.50 + 145

433.54 + 146

476.51 + 147

454.47 + 148

405.44 + 149

460.52 + 150

405.46 ++ 151

417.47 ++++ 152

420.50 + 153

406.45 + 154

473.50 + 155

455.57 + 156

457.54 ++ 157

442.45 + 158

421.48 + 159

474.54 + 160

401.47 +++ 161

456.43 + 162

442.52 + 163

361.41 + 164

421.48 + 165

450.45 + 166

453.51 + 167

395.43 + 168

342.37 + 169

358.37 + 170

411.43 + 171

445.50 + 172

414.47 + 173

421.48 + 174

402.46 + 175

493.48 +++ 176

375.44 ++ 177

420.50 +++ 178

426.48 +++ 179

404.43 +++ 180

416.49 + 181

491.55 +++ 182

439.48 + 183

396.41 + 184

454.50 185

428.50 + 186

461.59 +++ 187

445.53 ++++ 188

418.46 + 189

389.46 + 190

513.58 + 191

498.57 + 192

403.45 ++++ 193

491.55 ++ 194

500.55 + 195

455.50 + 196

441.46 + 197

342.36 +++ 198

432.51 +++ 199

460.61 +++ 200

410.44 + 201

403.49 ++ 202

401.52 +++ 203

404.43 ++++ 204

445.53 +++ 205

467.58 ++++ 206

466.59 ++++ 207

292.31 ++ 208

362.40 + 209

446.52 +++ 210

468.57 ++++ 211

427.47 +++ 212

386.42 +++ 213

341.38 +++ 214

427.47 ++++ 215

428.46 +++ 216

494.51 ++ 217

416.44 +++ 218

418.51 + 219

469.55 +++ 220

485.57 ++++ 222

457.54 +++ 224

416.49 ++ 225

430.52 ++ 226

481.56 +++ 227

495.63 + 228

473.60 +++ 229

535.58 230

343.35 +++ ++++ compound exhibited MAPKAPK5 IC₅₀ 1-100 nM +++ compoundexhibited MAPKAPK5 IC₅₀ 101-500 nM ++ compound exhibited MAPKAPK5 IC₅₀501-1000 nM + compound exhibited MAPKAPK5 IC₅₀ >1000 nM

EXAMPLE 2 Development of an Assay for the Identification of Regulatorsof the Expression of MMP1 by Activated Primary Synovial Fibroblasts

To identify compounds that decrease the ECM-degrading activity of cells,the ECM-degrading activity of cells may be induced to allow properdetection of this activity, and to achieve a clearer read-out. In thecontext of RA, the cells of choice are mammalian synovial fibroblastsand the triggers that may be used to induce the ECM-degrading activityare cytokines relevant in the field of arthritis: for instance TNF-α,IL1β, IL6, OSM, IL17, and MIF1-α. This list is not comprehensive due tothe plethora of cytokines potentially involved in the RA pathogenesis(Smolen and Steiner, 2003). To set up an in vitro assay that is as closeas possible to the complexity of the pathology, the trigger appliedshould be a mixture of factors generated by contactingcytokine-producing cells relevant in the field of arthritis, such asmonocytes, macrophages, T-cells, and B-cells, with a trigger. Thecytokine-producing cells will respond to the contact by producing acomplex and unbiased mixture of factors. If the cytokine-producing cellused is also found in a pannus, and the cytokine applied to produce thistrigger is found in the synovial fluid of rheumatoid arthritis patients,the mixture of factors ultimately produced will contain part of thefactors that are present in the joints of arthritis patients.

Principle of the ‘MMP Assay’

Matrix Metallo Proteases (MMPs) possess various physiological roles, ase.g. the maturation of other proteases, growth factors, and thedegradation of extra-cellular matrix components. MMP1 is one of themembers of the MMP family that is able to degrade native collagen, themain component of bone and cartilage. An increased expression of MMP1 bysynovial fibroblasts (SFs) is diagnostic for the progression of thearthritic disease and is predictive for erosive processes in the joint(Cunnane et al., 2001). The expression of MMP1 by SFs: can be increasedby the activation of SFs with triggers relevant for rheumatoidarthritis, as cytokines like TNF-α or IL1β (Andreakos et al., 2003).Taken together, measurement of the levels of MMP1 produced by activatedSFs is a readout that is highly relevant in the context of RA as thisevent reflects the level of activation of SFs towards an erosivephenotype as it is seen in the pannus. If a reduced expression of acandidate drug target in activated SFs leads to the reduction of MMP1expression by these cells, the drug target is then proven to be involvedin the regulation of MMP1 expression and thus considered relevant forthe development of therapeutic strategies for the treatment of RA.

In the following examples, the development of an assay, further referredto as ‘MMP assay’, monitors the MMP1 production by synovial fibroblasts(SFs) in response to diverse activating triggers (Example 2.1). The useof this assay is then described for the validation of gene products thatare considered drug targets for the development of RA therapies (Example2.2). The validation of drug targets is performed using recombinantadenoviruses, further referred to as knock-down viruses or Ad-siRNAs,that mediate the expression in cells of shRNA's which reduce theexpression levels of targeted genes by a RNAi (RNA interference)-basedmechanism, (see WO 03/020931). The identification of compoundsmodulating the activity of the validated drug targets is then describedin Table B. The use of the ‘MMP assay’ for the testing of compounds thatmodulate the activity of the drug targets identified is describedfurther below.

Control Viruses Used:

The control viruses used in these studies are listed below. dE1/dE2Aadenoviruses are generated from these adapter plasmids byco-transfection of the helper plasmid pWEAd5AflII-rITR.dE2A in PER.E2Apackaging cells, as described in W099/64582.

Negative Control Viruses.

-   Ad5-eGFP_KD: Target sequence: GCTGACCCTGAAGTTCATC (SEQ ID NO: 1).    Cloned using Sap1-sites into vector and virus generated as described    in WO03/02093 1.-   Ad5-Luc_v13_KD: Target sequence GGTTACCTAAGGGTGTGGC (SEQ ID NO: 2).    Cloned using Sap1-sites into vector and virus generated as described    in WO03/02093 1.-   Ad5-M6PR_v1_KD: Target sequence CTCTGAGTGCAGTGAAATC (SEQ ID NO: 3).    Cloned using Sap1-sites into vector and virus generated as described    in WO03/02093 1.    Positive Control Viruses:-   Ad5-MMP1_v10_KD: Target sequence ACAAGAGCAAGATGTGGAC (SEQ ID NO: 4).    Cloned using Sap1-sites into vector and virus generated as described    in WO03/02093 1.    Viruses Used for Target Validation:-   Ad5-MAPKAPK5_v13_KD: Target sequence CGGCACTTTACAGAGAAGC (SEQ ID NO:    5). Cloned using Sap1-sites into vector and virus generated as    described in WO03/020931.-   Ad5-MAPKAPK5_v12_KD: Target sequence ATGATGTGTGCCACACACC (SEQ ID NO:    6). Cloned using Sap1-sites into vector and virus generated as    described in WO03/02093 1.

EXAMPLE 2.1 Development of the MMP Assay

A 384-well format ELISA for measurement of MMP1 is developed. Variousprimary antibodies are tested, as well as various ELISA protocols. Thefollowing protocol is developed and validated to measure MMP1 levels inSF supernatant in 384 well plates: white Lumitrac 600 384 well plates(Greiner) are coated with 2 μg/mL anti-MMP1 antibody MAB1346 (Chemicon).The antibody is diluted in buffer 40 (1.21 g Tris base (Sigma), 0.58 gNaCl (Calbiochem) and 5 ml 10% NaN₃ (Sigma) in 1 L milliQ water andadjusted to pH 8.5). After overnight incubation at 4° C., plates arewashed with PBS (80 g NaCl, 2g KCl (Sigma), 11.5 g Na₂HPO₄.7H₂O and 2 gKH₂PO₄ in 10 L milliQ; pH 7.4) and blocked with 100 μL/well Caseinbuffer (2% Casein (VWR International) in PBS). Next day, casein bufferis removed from ELISA plates and replaced by 50 μL/well EC buffer (4 gcasein, 2.13 g Na₂HPO₄ (Sigma), 2 g bovine albumin (Sigma), 0.69 gNaH₂PO₄.H₂O (Sigma), 0.5 g CHAPS (Roche), 23.3 g NaCl, 4 mL 0.5 M EDTApH 8 (Invitrogen), 5 mL 10% NaN₃ in 1 L milliQ and adjusted to pH 7.0).0.25 mM DTT (Sigma) is added to the thawed samples plates. After removalof the EC buffer, 20 μL of sample is transferred to the ELISA plates.After overnight incubation at 4° C. plates are washed twice with PBS andonce with PBST (PBS with 0.05% Tween-20 (Sigma)) and incubated with 35μL/well biotinylated anti-MMP1 antibody solution (R&D). This secondaryantibody is diluted in buffer C (0.82 g NaH₂PO₄.H₂O, 4.82 g Na₂HPO₄,46.6 g NaCl, 20 g bovine albumin and 4 mL 0.5M EDTA pH 8 in 2 L milliQand adjusted to pH 7.0) at a concentration of 5 μg/mL. After 2 h ofincubation at RT, plates are washed as described above and incubatedwith 50 μL/well streptavidin-HRP conjugate (Biosource). Streptavidin-HRPconjugate is diluted in buffer C at a concentration of 0.25 μg/mL. After45 min, plates are washed as described above and incubated for 5 minwith 50 μL/well BM Chem ELISA Substrate (Roche). Readout is performed onthe Luminoscan Ascent Luminometer (Labsystems) with an integration timeof 200 msec or with an Envision reader (Perkin Elmer).

The increase of MMP1 expression by SFs upon treatment with cytokinesrelevant in the field of RA (TNF-α, IL1β and OSM) or a combinationthereof is shown in FIG. 2 as white bars. For this experiment, SFs areseeded in 96 well plates, 3,000 cells/well. 24 h later, the medium ischanged to M199 medium supplemented with 1% FBS. One day after themedium change, cytokines or combinations thereof are added to thecultures, each cytokine being added to a final concentration of 25ng/mL. 72 h after cytokine addition, the supernatant is collected andprocessed in the MMP1 ELISA as described in the protocol given above. Inparallel with this experiment, SFs are triggered, using the sameprotocol, with the supernatant of THP1 cells (2-fold diluted in M199+1%FBS) treated with the same cytokines or combinations of cytokines for 48h in M199 medium+1% FBS. MMP1 levels for these samples are shown in FIG.2 as grey bars. The induction of the MMP1 expression by SFs triggeredwith the supernatants of TNF-α-treated THP1 cells is stronger (>4.5 foldinduction) as compared to the SFs triggered with recombinant TNF-α alone(3-fold induction) and almost equals the 5-fold induction obtained by amixture of 3 purified cytokines (TNF-α, IL1βb, OSM). This resultindicates that the supernatant of TNF-α-induced THP1 cells contains,besides TNF-α, additional pro-inflammatory factors that activate SFstowards MMP1 expression. As the role of TNF-α in the RA pathogenesis isvalidated (TNF-α-blockers such as Infliximab and Etanercept show someefficacy in the treatment of RA patients) and the THP-1 cells arerepresentative for monocytes/macrophages present in the joint of RApatients, the TNF-α-based trigger mixture prepared by contacting THP-1cells with TNF-α will contain factors present in the joints of RApatients and subsequently is relevant to RA. This TNF-α-based complextrigger, further referred to as the ‘complex trigger’, will further beused as basis for the ‘MMP assay’.

Inhibition of the activation of SF by the ‘complex trigger’ is shownusing dexamethasone, a potent anti-inflammatory agent that also stronglyreduces collagen-induced arthritis in rodents (Yang et al., 2004) (FIG.3). Dexamethasone is shown to dose-dependently reduce amounts of MMP1produced by complex trigger activated SFs. SFs are seeded at a densityof 3000 cells/well in 96 well plates. 24 hrs after seeding, increasingconcentrations of dexamethasone are added to the cells. After overnightincubation, medium of every well is refreshed to supernatant of THP-1cells treated with TNF-α (50% diluted in M199+0.5% FBS), and the sameconcentration of dexamethasone as added the day before. 48 hrs aftertreatment, the supernatant is collected and subjected to the MMP1 ELISAdescribed above. The addition of dexamethasone clearly reduced the MMP1expression by SFs, with an IC₅₀ value of about 1 nM (see FIG. 3). Thesedata show that the MMP1 expression by activated SFs can be reduced bythe addition of a physiologically relevant inhibitor and represent aproof of principle for the ‘MMP assay’.

EXAMPLE 2.2 MAPKAPK5 Modulates SF ‘Complex Trigger’-induced MMP1Expression

(A) Ad-siRNA Virus Functions to Knock Down MAPKAPK5 Expression.

Recombinant adenoviruses mediating the expression of siRNA's targetingMAPKAPK5 and eGFP are generated according to the procedure described inWO03/02093 1. The target sequence used in the recombinant adenovirus is:CGGCACTTTACAGAGAAGC (SEQ ID NO: 5) as well as ATGATGTGTGCCACACACC (SEQID NO: 6). The target sequence within the eGFP mRNA used in therecombinant adenovirus is: GCTGACCCTGAAGTTCATC (SEQ ID NO: 1). Thesesequences are cloned into the adapter plasmid using Sapl sites. dE1/dE2Aadenoviruses are generated from these adapter plasmids byco-transfection of the helper plasmid pWEAd5AflII-rITR.dE2A in PER.E2Apackaging cells, as described in WO99/64582.

The functionality of an adenovirus targeting MAPKAPK5 is tested asfollows. These adenoviruses are used to infect primary human SFscultured in petri dishes as follows. On day 1, 500.000 SFs are seededper petri dish. One day later, the cells are infected withAd5-MAPKAPK5-v13_KD (1.6E9 VP/mL) or Ad5-eGFP-v5_KD (1.3E10 VP/mL) at anMOI of 4000 (based on the titers (number of virus particles per mL)defined for the viruses by Q-rt-PCR). On day 7, cells are detached fromthe petri dish according to standard procedure using a trypsin EDTAsolution. The trypsin is then neutralized by addition of DMEM growthmedium supplemented with 10% FBS. The cells are then collected by acentrifugation step (1000 rpm, 5 min). The pellet is lysed in 100 μL offresh RIPA buffer (50 mM Tris pH7.5, 150 mM NaCl, 1% deoxycholate, 1%Triton X100, 0.1% SDS). The samples are then sonicated for 10 sec. Theprotein concentration of the samples is then determined using the BCAkit (Pierce, Cat N° 23227) as described by the provider, using BSA as astandard. To 30 μg of cell lysate diluted to 19.5 μl in RIPA buffer, 3.5μL of reducing agent (NuPage reducing agent N° 10, Invitrogen NP0004)and 7.5 μL of sample buffer (NuPage LDS sample buffer, InvitrogenNP0007) are added. The 30 μL sample is then boiled for 5 min and loadedon a 10% polyacrylamide gel (Invitrogen NP0301). To allow the estimationof the level of protein knock-down, 15 μg, 7.5 μg and 3.75 μg of thelysate of the Ad5-eGFP-v5_KD infected cells are also loaded onto thegel. The gel is then run for 2 hours at 100V in 1× MOPS/SDS NuPagerunning buffer (Invitrogen NP001). 10 μl of Seablue Plus Prestainedstandard (Invitrogen LC5925) is used to estimate protein size on thegel. The proteins on the gel are then transferred onto a PVDF membrane(Invitrogen LC2002) by a wet blotting procedure using a transfer bufferprepared by mixing 100 ml Nupage Transfer buffer 20*(NP0006-1), 400 mLmethanol and 1500 mL Milli Q water. Before the transfer, the membrane isfirst soaked in methanol and in transfer buffer. The transfer isperformed at 100V for 90 minutes. The membrane is then blocked by 30 minsoaking in blocking buffer (2% blocking blocking powder (Amersham, RPN2109) prepared in PBST (PBS supplemented with 0.1% Tween 20 (Sigma,P1379)). After blocking, the immunodetection is performed using a mousemonoclonal antibody against MAPKAPK5 (BD Biosciences, Cat N° 612080)diluted 250 fold in blocking buffer. After overnight incubation withthis primary antibody, the membrane is washed 3 times with PBST andincubated 1 hr with the secondary antibody ((Polyclonal goat anti-mouseIg, HRP conjugated (DAKO P0447) diluted 50000 fold in blocking buffer.The blot is then washed 3 times in PBST and the detection is performedwith ECL advance (RPN2109, Amersham) on a Kodakimager according to themanufacturers instructions. The Western Blotting revealed a lowerexpression level of MAPKAPK5 in the Ad5-MAPKAPK5-v13_KD infected cellscompared to the cells infected with the Ad5-eGFP-v5_KD negative controlvirus. Comparison with the diluted Ad5-eGFP-v5_KD infected samplesallowed to estimate the reduction in expression to be 2-fold. Equalloading of the 30 μg samples is demonstrated by immunodetection ofβ-actin after removal of the MAPKAPK5 antibody by a ‘strippingprocedure’ (5 minutes boiling of the membrane in PBST). Immunodetectionof β-actin is performed according to the method described for MAPKAPK5detection, but using a goat polyclonal antibody against β-actin (SantaCruz, Cat N° SC-1615) at a 1000 fold dilution as primary antibody and arabbit anti goat antibody at a 50000 fold dilution as a secondaryantibody. Results of this experiment are given in FIG. 4. Takentogether, this experiment demonstrated the functionality of the Ad-siRNAvirus produced to reduce the MAPKAPK5 expression levels in primary humanSFs.

(B) MAPKAPK5 Knock-Down Ad-siRNA Reduces SF-Induced MMP1 Expression

The efficacy of Ad5-MAPKAPK5-v13_KD virus in the ‘MMP assay’ is testedas follows. Day 1, SFs (passage 9 to 10) are seeded in 96 well plates ata density of 3000 cells per well in complete synovial growth medium(Cell Applications). One day later, the cells are infected withincreasing amounts (3, 6; 9, 12 or 15 l) of following viruses:Ad5-eGFP-v5_KD, Ad5-MAPKAPK5-v12_KD, Ad5-MAPKAPK5-v13_KD,Ad5-MMP1-v10_KD. The virus load is corrected by addition of the neutralvirus Ad5-Luc-v13_KD to bring the final virus volume on the cells to 15μL in every well. This correction guarantees that the effects observeddo not result from the virus load applied to the cells. The cells arethen incubated for 5 days before the activation step. This step involvesthe replacement, in every well, of the growth medium by 75 μL of M199medium supplemented with 25 μL of ‘complex trigger’. 48 hrs after theactivation step, the supernatant is collected and subjected to the MMP1ELISA as described in Example 1. The results of the experiment are shownin FIG. 5. The quality of the experiment is demonstrated by the efficacyof the Ad-siRNA virus targeting MMP1 itself. This positive control virusstrongly reduces the MMP1 expression by SFs, whereas the negativecontrol virus, designed to target the expression of luciferase, does notinfluence the levels of MMP1 expression. Two viruses used to validatethe MAPKAPK5 target (Ad5-MAPKAPK5-v12_KD and Ad5-MAPKAPK5-v13) do alsolead to a clear reduction of the complex trigger induced MMP1 expressionby primary human SFs. It can be concluded, from this experiment, thatMAPKAPK5 represents a valuable drug target that is shown to modulateMMP1 expression in SFs. Similarly, the inhibition of MAPKAPK5 enzymaticactivity by a small molecule compound is expected to reduce the ‘complexcytokine’ induced MMP1 expression in the ‘MMP assay’. The inhibition ofMAPKAPK5 enzymatic activity by a small molecule compound is alsopredicted to reduce the degradation of the joint associated with RA.

(C) In vitro ‘MMP Assay’ Testing of Compounds Inhibiting MAPKAPK5

Compounds inhibiting the MAPKAPK5 activity in a biochemical assay (i.e.cell free, using purified enzyme), are tested in the ‘MMP assay’according to following protocol.

The compound master stocks (all at 10 mM concentration in 100% DMSO) arediluted 10-fold in water (Distilled water, GIBCO, DNAse and RNAse free)to obtain a 1 mM intermediate work stock in 10% DMSO. This intermediatework stock is further diluted either 3-fold (or 10-fold) in 10% DMSO toobtain an intermediate work stock of 333 μM (or 100 μM) concentration,respectively, in 10% DMSO. The 1 mM as well as 333 μM (or 100 μM)intermediate work stocks are then further diluted 10-fold in 1.1% DMSOto obtain the 10× workstocks at 100 μM and 33.3 μM (or 10 μM)concentration in 2% DMSO. This 10× work stock is then diluted 10-fold inM199 medium supplemented with 1% FBS to obtain the final ‘1 compoundpreparation’ containing the compounds at 10 μM and 3.33 μM (or 1 μM) aswell as 0.2% DMSO. These are the final conditions at which the compoundsare tested on the cells. In parallel, the 10× work stock is diluted10-fold in ‘complex trigger’ (i.e. the supernatant of TNF-α treated THP1cells produced as described in Example 1) that is diluted 2-fold in M199supplemented with 1% FBS to produce the ‘1× compound in 50% complextrigger preparation’.

At day 1, RASFs are seeded in 96 well plates (Flat bottom, tissueculture treated, Greiner) at a density of 3000 cells/well in completesynovial growth medium (Cell Applications). Day 5, the compounds areadded to the cultured cells as follows. Medium is completely removedfrom the cells and replaced by 75 μL of the ‘1× compound preparations’containing the compounds at either 10 μM or 3.33 μM (or 1 μM) in M199medium supplemented with 1% FBS and 0.2% DMSO. After an incubationperiod of 2 hours, which allows the compounds to equilibrate and enterthe cells, 25 μL of the ‘1× compound in 50% complex triggerpreparations’ are added to the wells on top of the ‘1× compoundpreparation’, in the wells containing the corresponding compounds atcorresponding concentration. In this way, an 8-fold diluted complextrigger is ultimately applied to the cells. An incubation of 48 hrs isthen performed and 20 μl of the cell supernatant is then processed inthe MMP1 ELISA as described above, delivering raw data (RLU: relativeluminescence units). Following controls are included in the experiments.A maximal signal control, in which the cells are activated by thecomplex trigger but only the 0.2% DMSO vehicle (and thus no compound) isadded. This control indicates the maximal level of MMP1 that can beachieved in the test. A minimal signal control is also included in theseexperiments. Here, cells are not triggered. The medium of the cells isthen changed to 100 μl M199 medium supplemented with 1% FBS at day 5.This control returns the basal MMP1 levels produced by the RASFs. Thepercent inhibition of the MMP1 expression achieved by the compounds isthen calculated based on the RLU data returned by the ELISA withfollowing formula: [[(maximal MMP1 levels−minimal MMP1 levels)−(MMP1level compound X at concentration Y−minimal MMP1 levels)]/(maximal MMP1levels−minimal MMP1 levels)]×100.

Toxicity of the compounds is assessed as follows. Day 1, SFs are seededin white, tissue culture treated 96 well plates at a density of 3000cells per well in 100 μL complete synovial growth medium. The compoundhandling, compound addition to the cells as well as activation of thecells is further performed as described above in this example for thedetermination of the MMP I levels. After the 48 hrs incubation period,the medium is removed from the wells, replaced by 50 μL fresh M199medium supplemented with 1% FBS. 50 μL of substrate (Promega CelltiterGlow cell viability kit) is then added to the wells. After an incubationperiod of 10 min, luminescence signal is measured. A reduction of theluminescence signal by more than 50% as compared to the maximal controlwells is considered to reflect significant toxicity. No toxicity isobserved for the compounds tested in the ‘MMP assay’.

It should be understood that factors such as the differential cellpenetration capacity of the various compounds can contribute todiscrepancies between the activity of the compounds in the in vitrobiochemical and cellular MMP assays.

For the purpose of Table 2 below, MMP1 EC₅₀ of each compound, which canbe determined using the assay method described herein, is expressed asfollows:

TABLE 2 MMP1 Compd EC₅₀ ID (nM) 1 * 2 * 3 4 *** 5 *** 6 * 7 * 8 9 * 10 *11 * 12 * 13 ** 14 * 15 * 16 * 17 * 18 * 19 ** 20 * 21 * 22 * 23 * 24 *25 * 26 ** 27 * 28 ** 29 * 30 * 31 * 32 * 33 * 34 ** 35 * 36 * 37 * 38 *39 * 40 * 41 * 42 * 43 * 44 * 45 * 46 * 47 * 48 * 49 * 50 * 51 * 52 *53 * 54 * 55 * 56 * 57 * 58 * 59 * 60 * 61 * 62 * 63 * 64 * 65 * 66 *67 * 68 * 69 * 70 * 71 * 72 * 73 * 74 * 75 * 76 * 77 * 78 * 79 * 80 *81 * 82 * 83 * 84 * 85 ** 86 * 87 * 88 * 89 ** 90 *** 91 * 92 * 93 *94 * 95 *** 96 * 97 * 98 ** 99 * 100 * 101 * 102 * 103 * 104 * 105 *106 * 107 * 108 *** 109 * 111 * 112 *** 114 * 115 * 116 * 117 * 118 *119 * 120 * 121 * 122 *** 123 *** 124 * 125 * 126 * 127 *** 128 * 129 *130 * 131 * 132 * 133 *** 134 *** 135 * 136 * 137 *** 138 * 139 * 140 *141 ** 142 ** 143 ** 144 *** 145 ** 146 ** 147 *** 148 ** 149 * 150 *151 *** 152 *** 153 *** 154 **** 155 *** 156 *** 157 * 158 * 159 ***160 * 161 * 162 ** 163 * 164 * 165 * 166 * 167 * 168 * 169 *** 170 ***171 *** 172 * 173 * 174 *** 175 ** 176 * 177 *** 178 *** 179 *** 180181 * 182 * 183 *** 184 *** 185 ** 186 ** 187 *** 188 ** 189 ** 190 ***191 * 192 ** 193 ** 194 ** 195 * 196 * 197 ** 198 *** 199 *** 200 ***201 * 202 * 203 ** 204 ** 205 * 206 * 207 * 208 * 209 * 210 ** 211 * 212*** 213 *** 214 * 215 * 216 * 217 * 218 * 219 ** 220 * 222 ** 224 *225 * 226 * 227 * 228

EXAMPLE 3 Assay to Assess Effect of Compounds on Cytokine Release byHuman PBMCs

Human peripheral blood mononuclear cells (PBMCs) are isolated from“buffy coats” prepared from the blood of healthy volunteers, isolatedessentially according to method of Boyum (1984). In brief, buffy coat isdiluted 1:1 with 1× PBS (Gibco) and 30 mL is carefully put on top of 20mL Lymphoprep™ (Lucron Bioproducts) in 50 mL Falcon tubes. Aftercentrifugation (35 min 400 g, 18° C.) the mononuclear cells arecollected from the white interphase and washed 3 times with 1× PBS byresuspending and centrifugation (10 min, 200 g). Isolated PBMCs arefinally resuspended in RPMI 1640 (Cat. No. 21875, Gibco) that issupplemented with 10% heat-inactivated FBS (Hyclone).

For the assay PBMCs are seeded at 2.5E6 cells/mL in 160 μL in 96-wellplates (Nunc). Serial dilution of the test compounds are made first inDMSO (Sigma) and then diluted 50-fold in M199 medium (Gibco) containing1% heat-inactivated FBS. Compounds are further 1/10 diluted in the assayplates to obtain final DMSO concentration of 0.2%. Cells arepreincubated with the compounds for 1 hr at 37° C., 5% CO₂. Then, cellsare stimulated with LPS (Escherichia coli serotype 026:B6, Cat. No.L2654, Sigma) that is added in a volume of 20 μL to a finalconcentration of 1 μg/mL and cells are further cultured for 24 hr. Theplates are centrifuged and the supernatant is collected and stored at−80° C. until analysis of appropriate dilutions in ELISAs.

The following 384-well chemiluminescent ELISA protocol was developed tomeasure TNFα levels in the supernatant : White Lumitrac 600 384-wellplates (Greiner) are coated with (40 μL/well) anti-TNFα capture antibody(Cat. No. 551220, BD Pharmingen) that is diluted to 1 μg/mL in 1× PBS(Gibco). After overnight incubation at 4° C., plates are washed with 1×PBS (80 g NaCl, 2 g KCl (Sigma), 11.5 g Na₂HPO₄.7H2O and 2 g KH₂PO₄ in10 L milliQ; pH 7.4) and blocked with 100 μL/well buffer B (1× PBScontaining 1% BSA (Sigma), 5% sucrose (Sigma) and 0.05% NaN₃ (Sigma)).After 4 hr incubation at RT, blocking buffer is removed and plates arewashed once with PBST (lx PBS with 0.05% Tween-20 (Sigma)). Then, 40 μLof sample is transferred to the ELISA plates and plates are incubated at4° C. Next day, plates are washed 3 times (twice with PBST and once withPBS) and 35 μL/well biotinylated anti-TNFα antibody (Cat. No. 554511, BDPharmingen) diluted first to a concentration of 250 ng/ml in buffer D(1× PBS with 1% BSA) is added. After 2 h of incubation at RT, plates arewashed as described above and 35 μL/well of a 1/2000 dilution ofstreptavidin-HRP conjugate (Cat. No. SNN2004, Biosource) in buffer D isadded. After 45 min, plates are washed as described above and incubatedfor 5 min with 50 μL/well BM Chemiluminescence ELISA Substrate POD(Roche). Readout is performed on the Luminoscan Ascent Luminometer(Labsystems) with an integration time of 100 msec delivering raw data(RLU: relative luminescence units). The following controls are includedin the experiments, a maximal signal control, in which the cells areactivated by LPS but only the 0.2% DMSO vehicle (and thus no compound)is added. This control indicates the maximal level of TNFα that can beachieved in the test. A minimal signal control is also included in theseexperiments. Here, cells are not triggered. This control returns thebasal TNFα levels produced by the PBMCs. The percent inhibition (PIN) ofthe TNFα release, achieved by the compounds is then calculated based onthe RLU data returned by the ELISA with following formula: 100−[((TNFαlevel compound X at concentration Y−minimal TNFα levels)/(maximal TNFαlevels−minimal TNFα levels))×100]. Where compounds are tested at 8concentrations (⅓ serial dilution), EC50-values can be calculated bycurve fitting of the means of the PIN data achieved for a compound ateach test concentration.

To assay the effect of compounds on the release of IL1 and IL6 by LPSstimulated PBMC cultures, appropriate dilutions of the supernatant canbe measured using the same ELISA protocol as described above. Matchedpair antibodies for IL1 and IL6 ELISA (all from R&D Systems) may be usedas follows : anti-IL1 capture antibody (Cat. No. MAB601) used at 0.5μg/mL , biotinylated anti-IL1 detection antibody (Cat. No. BAF201) usedat 50 ng/mL ; anti-IL6 capture antibody (Cat. No. MAB206) used at 1μg/mL; biotinylated anti-IL6 detection antibody (Cat. No. BAF206) usedat 50 ng/mL.

For the purpose of Table 3 below, PBMC EC₅₀ of each compound, which canbe determined using the assay method described herein, is expressed asfollows:

TABLE 3 PBMC EC₅₀ Compound # (nM) 1 # 11 # 13 # 14 # 26 # 32 # 35 ## 36# 41 # 45 # 53 # 55 # 56 # 64 # 69 ## 71 # 82 # 83 # 85 # 89 # 90 # 93 #98 # 102 # 114 # 116 # 118 # 119 # 120 # 122 # 123 # 124 ## 125 # 127 #128 # 129 # 136 ### 137 # 138 ### 139 # 140 # 141 # 142 # 143 # 144 ####145 # 146 # 149 # 150 # 151 # 152 ### 153 #### 155 # 156 # 158 ### 159 #160 # 163 # 165 # 166 # 167 #### 168 # 169 # 171 # 173 # 176 # 177 # 178#### 179 # 181 # 186 # 187 # 188 # 192 # 193 # 197 # 198 # 203 # 204 #205 # 210 # 218 #

The present invention relates also to a method of treatment orprevention of inflammatory diseases, which comprises administering to asubject in need thereof, a therapeutically effective inhibitor ofMitogen-Activated Protein Kinase-Activated Protein Kinase 5 inhibitingamount of a compound according to Formula 1.

Another aspect of the present method invention relates to a method oftreatment or prophylaxis of a condition characterised by abnormal matrixmetallo proteinase activity, which comprises administering atherapeutically effective amount of a matrix metallo proteinaseinhibiting compound according to Formula 1.

A further aspect of the present method invention is a method oftreatment or prophylaxis of a condition selected from diseases involvingdegradation of extra-cellular matrix, which comprises administering atherapeutically effective matrix metallo proteinase inhibiting amount ofa compound according to Formula 1.

A yet further aspect of the present method invention is a method oftreatment or prophylaxis of a condition selected from diseases involvingabnormal cellular expression of MMP1, which comprises administering atherapeutically effective matrix metallo proteinase inhibiting amount ofa compound according to Formula 1.

A special embodiment of the present method invention is a method oftreatment or prevention of rheumatoid arthritis, which comprisesadministering to a subject in need thereof, a therapeutically effectiveamount of a compound according to Formula 1.

This invention also relates to the use of the present compounds in themanufacture of a medicament for treatment or prophylaxis of a conditionprevented, ameliorated or eliminated by administration of an inhibitorof Mitogen-Activated Protein Kinase-Activated Protein Kinase 5, or acondition characterised by abnormal collagenase activity, or a conditionselected from diseases involving inflammation, most preferably in forthe treatment of rheumatoid arthritis.

Administering of the compound of the present invention to the subjectpatient includes both self-administration and administration by anotherperson. The patient may be in need of treatment for an existing diseaseor medical condition, or may desire prophylactic treatment to prevent orreduce the risk for diseases and medical conditions affected by adisturbance in bone metabolism. The compound of the present inventionmay be delivered to the subject patient orally, transdermally, viainhalation, injection, nasally, rectally or via a sustained releaseformulation.

A preferred regimen of the present method comprises the administrationto a subject in suffering from a disease condition characterized byinflammatory, with an effective matrix metallo-protease inhibitingamount of a compound of the present invention for a period of timesufficient to reduce the abnormal levels of extracellular matrixdegradation in the patient, and preferably terminate, theself-perpetuating processes responsible for said degradation. A specialembodiment of the method comprises administering of an effective matrixmetallo-protease inhibiting amount of a compound of the presentinvention to a subject patient suffering from or susceptible to thedevelopment of rheumatoid arthritis, for a period of time sufficient toreduce or prevent, respectively, collagen and bone degradation in thejoints of said patient, and preferably terminate, the self-perpetuatingprocesses responsible for said degradation.

Toxicity and therapeutic efficacy of such compounds can be determined bystandard pharmaceutical procedures in cell cultures or experimentalanimals, e.g., for determining the LD50 (the dose lethal to 50% of thepopulation) and the ED50 (the dose therapeutically effective in 50% ofthe population). The dose ratio between toxic and therapeutic effects isthe therapeutic index and it can be expressed as the ratio LD50/ED50.Compounds that exhibit large therapeutic indices are preferred. Whilecompounds that exhibit toxic side effects may be used, care should betaken to design a delivery system that targets such compounds to thesite of affected tissue in order to minimize potential damage touninfected cells and, thereby, reduce side effects.

The data obtained from the cell culture assays and animal studies can beused in formulating a range of dosage for use in humans. The dosage ofsuch compounds lies preferably within a range of circulatingconcentrations that include the ED50 with little or no toxicity. Thedosage may vary within this range depending upon the dosage formemployed and the route of administration utilized. For any compound usedin the method of the invention, the therapeutically effective dose canbe estimated initially from cell culture assays. A dose may beformulated in animal models to achieve a circulating plasmaconcentration range that includes the IC50 (i.e., the concentration ofthe test compound which achieves a half-maximal inhibition of symptoms)as determined in cell culture. Such information can be used to moreaccurately determine useful doses in humans. Levels in plasma may bemeasured, for example, by high performance liquid chromatography.

A preferred therapeutically effective amount of the compound of thepresent invention to administer to a subject patient is about 0.1 mg/kgto about 10 mg/kg administered from once to three times a day. Forexample, an effective regimen of the present method may administer about5 mg to about 1000 mg of said compound of the present invention fromonce to three times a day. It will be understood, however, that thespecific dose level for any particular subject patient will depend upona variety of factors including the age, body weight, general health,sex, diet, time of administration, route of administration, rate ofexcretion, drug combination and the severity of the particularinflammatory condition. A consideration of these factors is well withinthe purview of the ordinarily skilled clinician for the purpose ofdetermining the therapeutically effective or prophylactically effectivedosage amount needed to prevent, counter, or arrest the progress of thecondition.

Compounds of the invention can be incorporated into pharmaceuticalcompositions suitable for administration. Such compositions typicallycomprise at least one compound of the invention and at least onepharmaceutically acceptable carrier. As used herein the language“pharmaceutically acceptable carrier” is intended to include solidcarriers such as lactose, magnesium stearate, terra alba, sucrose, talc,stearic acid, gelatin, agar, pectin, acacia or the like; and liquidssuch as vegetable oils, arachis oil and sterile water, or the like, anyand all solvents, dispersion media, coatings, antibacterial andantifungal agents, isotonic and absorption delaying agents, and thelike, compatible with pharmaceutical administration. This listing ofpharmaceutically acceptable carriers is not to be construed as limiting.The use of such media and agents for pharmaceutically active substancesis well known in the art. Except insofar as any conventional media oragent is incompatible with the active compound, use thereof in thecompositions is contemplated. Supplementary active compounds can also beincorporated into the compositions.

A pharmaceutical composition of the invention is formulated to becompatible with its intended route of administration. Examples of routesof administration include parenteral, e.g., intravenous, intradermal,subcutaneous, oral (e.g., inhalation), transdermal (topical),transmucosal, and rectal administration. Solutions or suspensions usedfor parenteral, intradermal, or subcutaneous application can include thefollowing components: a sterile diluent such as water for injection,saline solution, fixed oils, polyethylene glycols, glycerine, propyleneglycol or other synthetic solvents; antibacterial agents such as benzylalcohol or methyl parabens; antioxidants such as ascorbic acid or sodiumbisulfite; chelating agents such as ethylenediaminetetraacetic acid;buffers such as acetates, citrates or phosphates and agents for theadjustment of tonicity such as sodium chloride or dextrose. The pH canbe adjusted with acids or bases, such as hydrochloric acid or sodiumhydroxide. The parenteral preparation can be enclosed in ampoules,disposable syringes or multiple dose vials made of glass or plastic.

Pharmaceutical compositions suitable for injectable use include sterileaqueous solutions (where water soluble) or dispersions and sterilepowders for the extemporaneous preparation of sterile injectablesolutions or dispersion. For intravenous administration, suitablecarriers include physiological saline, bacteriostatic water, CremophorELTM (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In allcases, the composition must be sterile and should be fluid to the extentthat easy syringability exists. It must be stable under the conditionsof manufacture and storage and must be preserved against thecontaminating action of microorganisms such as bacteria and fungi. Thecarrier can be a solvent or dispersion medium containing, for example,water, ethanol, polyol (for example, glycerol, propylene glycol, andliquid polyetheylene glycol, and the like), and suitable mixturesthereof. The proper fluidity can be maintained, for example, by the useof a coating such as lecithin, by the maintenance of the requiredparticle size in the case of dispersion and by the use of surfactants.Prevention of the action of microorganisms can be achieved by variousantibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In manycases, it will be preferable to include isotonic agents, for example,sugars, polyalcohols such as manitol, sorbitol, sodium chloride in thecomposition. Prolonged absorption of the injectable compositions can bebrought about by including in the composition an agent which delaysabsorption, for example, aluminum mono stearate and gelatin.

Sterile injectable solutions can be prepared by incorporating the activecompound (e.g., a compound according to an embodiment of the invention)in the required amount in an appropriate solvent with one or acombination of ingredients enumerated above, as required, followed byfiltered sterilization. Generally, dispersions are prepared byincorporating the active compound into a sterile vehicle which containsa basic dispersion medium and the required other ingredients from thoseenumerated above. In the case of sterile powders for the preparation ofsterile injectable solutions, the preferred methods of preparation arevacuum drying and freeze-drying which yields a powder of the activeingredient plus any additional desired ingredient from a previouslysterile-filtered solution thereof.

Oral compositions generally include an inert diluent or an ediblecarrier. They can be enclosed in gelatin capsules or compressed intotablets. For the purpose of oral therapeutic administration, the activecompound can be incorporated with excipients and used in the form oftablets, troches, or capsules. Oral compositions can also be preparedusing a fluid carrier for use as a mouthwash, wherein the compound inthe fluid carrier is applied orally and swished and expectorated orswallowed.

Pharmaceutically compatible binding agents, and/or adjuvant materialscan be included as part of the composition. The tablets, pills,capsules, troches and the like can contain any of the followingingredients, or compounds of a similar nature: a binder such asmicrocrystalline cellulose, gum tragacanth or gelatin; an excipient suchas starch or lactose, a disintegrating agent such as alginic acid,Primogel, or corn starch; a lubricant such as magnesium stearate orSterotes; a glidant such as colloidal silicon dioxide; a sweeteningagent such as sucrose or saccharin; or a flavoring agent such aspeppermint, methyl salicylate, or orange flavoring.

For administration by inhalation, the compounds are delivered in theform of an aerosol spray from pressured container or dispenser whichcontains a suitable propellant, e.g., a gas such as carbon dioxide, or anebulizer.

Systemic administration can also be by transmucosal or transdermalmeans. For transmucosal or transdermal administration, penetrantsappropriate to the barrier to be permeated are used in the formulation.Such penetrants are generally known in the art, and include, forexample, for transmucosal administration, detergents, bile salts, andfusidic acid derivatives. Transmucosal administration can beaccomplished through the use of nasal sprays or suppositories. Fortransdermal administration, the active compounds are formulated intoointments, salves, gels, or creams as generally known in the art.

The compounds can also be prepared in the form of suppositories (e.g.,with conventional suppository bases such as cocoa butter and otherglycerides) or retention enemas for rectal delivery.

In one embodiment, the active compounds are prepared with carriers thatwill protect the compound against rapid elimination from the body, suchas a controlled release formulation, including implants andmicroencapsulated delivery systems. Biodegradable, biocompatiblepolymers can be used, such as ethylene vinyl acetate, polyanhydrides,polyglycolic acid, collagen, polyorthoesters, and polylactic acid.Methods for preparation of such formulations will be apparent to thoseskilled in the art. The materials can also be obtained commercially fromAlza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions(including liposomes targeted to infected cells with monoclonalantibodies to viral antigens) can also be used as pharmaceuticallyacceptable carriers. These can be prepared according to methods known tothose skilled in the art.

It is especially advantageous to formulate oral or parenteralcompositions in dosage unit form for ease of administration anduniformity of dosage. Dosage unit form as used herein refers tophysically discrete units suited as unitary dosages for the subject tobe treated; each unit containing a predetermined quantity of activecompound calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical carrier. The specificationfor the dosage unit forms of the invention are dictated by and directlydependent on the unique characteristics of the active compound and theparticular therapeutic effect to be achieved, and the limitationsinherent in the art of compounding such an active compound for thetreatment of individuals.

The pharmaceutical compositions can be included in a container, pack, ordispenser together with instructions for administration.

A compound according to an embodiment of the invention may be providedas a salt, preferably as a pharmaceutically acceptable salt of compoundsof formula 1. Examples of pharmaceutically acceptable salts of thesecompounds include those derived from organic acids such as acetic acid,malic acid, tartaric acid, citric acid, lactic acid, oxalic acid,succinic acid, fumaric acid, maleic acid, benzoic acid, salicylic acid,phenylacetic acid, mandelic acid, methanesulphonic acid,benzenesulphonic acid and p-toluenesulphonic acid, mineral acids such ashydrochloric and sulphuric acid and the like, giving methanesulphonate,benzenesulphonate, p-toluenesulphonate, hydrochloride and sulphate, andthe like, respectively or those derived from bases such as organic andinorganic bases. Examples of suitable inorganic bases for the formationof salts of compounds for this invention include the hydroxides,carbonates, and bicarbonates of ammonia, lithium, sodium, calcium,potassium, aluminum, iron, magnesium, zinc and the like. Salts can alsobe formed with suitable organic bases. Such bases suitable for theformation of pharmaceutically acceptable base addition salts withcompounds of the present invention include organic bases which arenontoxic and strong enough to form salts. Such organic bases are alreadywell known in the art and may include amino acids such as arginine andlysine, mono-, di-, or trihydroxyalkylamines such as mono-, di-, andtriethanolamine, choline, mono-, di-, and trialkylamines, such asmethylamine, dimethylamine, and trimethylamine, guanidine;N-methylglucosamine; N-methylpiperazine; morpholine; ethylenediamine;N-benzylphenethylamine; tris(hydroxymethyl) aminomethane; and the like.

Salts of compounds according to an embodiment of the invention may beprepared in a conventional manner using methods well known in the art.Acid addition salts of said basic compounds may be prepared bydissolving the free base compounds according to the first or secondaspects of the invention in aqueous or aqueous alcohol solution or othersuitable solvents containing the required acid. Where a compound of theinvention contains an acidic function, a base salt of said compound maybe prepared by reacting said compound with a suitable base. The acid orbase salt may separate directly or can be obtained by concentrating thesolution e.g. by evaporation. The compounds of this invention may alsoexist in solvated or hydrated forms.

It will be appreciated by those skilled in the art that the foregoingdescription is exemplary and explanatory in nature, and is intended toillustrate the invention and its preferred embodiments. Through routineexperimentation, an artisan will recognise apparent modifications andvariations that may be made without departing from the spirit of theinvention. Thus, the invention is intended to be defined not by theabove description, but by the following claims and their equivalents.

References

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From the foregoing description, various modifications and changes in thecompositions and methods of this invention will occur to those skilledin the art. All such modifications coming within the scope of theappended claims are intended to be included therein.

It should be understood that factors such as the differential cellpenetration capacity of the various compounds can contribute todiscrepancies between the activity of the compounds in the in vitrobiochemical and cellular assays.

All publications, including but not limited to patents and patentapplications, cited in this specification are herein incorporated byreference as if each individual publication were specifically andindividually indicated to be incorporated by reference herein as thoughfully set forth.

The chemical names of compounds of invention given in this applicationare generated using MDL's ISIS Draw Autonom Software tool and are notverified. Preferably, in the event of inconsistency, the depictedstructure governs.

1. A compound according to formula IVa, IVb, or IVc:

wherein L is —CON(H)—CH₂—CH₂—; the ring P is substituted orunsubstituted heterocycloalkyl; the subscript n, is selected from 1-4;each R^(8a) is independently selected from hydrogen, substituted orunsubstituted alkyl, alkoxy, cyano, and halo; and R⁹ is independentlyselected from substituted or unsubstituted aryl and heteroaryl; or apharmaceutically acceptable salt, thereof; or stereoisomers or tautomersthereof.
 2. A compound according to formula IVa, IVb, or IVc:

wherein L is —OCH₂—CH₂— or —NHCH₂—CH₂—; the ring P is substituted orunsubstituted heterocycloalkyl; the subscript n, is selected from 1-4;each R^(8a) is independently selected from hydrogen, substituted orunsubstituted alkyl, alkoxy, cyano, and halo; and R⁹ is independentlyselected from substituted or unsubstituted aryl and heteroaryl; or apharmaceutically acceptable salt, thereof; or stereoisomers or tautomersthereof.
 3. A compound according to any one of claims 1 and 2, whereinthe ring P is substituted or unsubstituted piperidine, morpholine orpiperazine.
 4. A compound according to formula III;

wherein R¹ is H, or substituted or unsubstituted alkyl; R² is H, loweralkyl, lower cycloalkyl and lower alkyl-lower cycloalkyl, optionallysubstituted with one or more of F and Cl; R⁸ is

L is selected from a single bond, alkylene, —O—, —N(R^(8e))—,—CO—,—CO₂—, —SO—, —SO₂—, —CON(R^(8e))—, —SO₂N(R^(8e))—, —N(R^(8e))CO—,—N(R^(8e))SO₂—, —N(R^(8e))CO N(R^(8e))—, —N(R^(8e))SO₂ N(R^(8e))—;—O(CR^(8e) ₂)_(m1)—, or —CON(R^(8e))(CR^(8e) ₂)_(m1)—; each R^(8a)independently selected from hydrogen, substituted or unsubstitutedalkyl, alkoxy, cyano, and halo; R^(8d) is selected from substituted orunsubstituted heterocycloalkyl; R^(8e) is selected from H, substitutedor unsubstituted alkyl and substituted or unsubstituted cycloalkyl; thesubscript m1 is 1, 2, 3, or 4; the subscript n is 1, 2, 3, or 4; andwherein R⁹ is

and each of A¹, A² and A³ is independently selected from S, O, N,NR^(9a), and CR^(9a); each of R^(9a)is independently H or substituted orunsubstituted alkyl; and R^(9b) is CONH₂, CONHMe, or CN; or apharmaceutically acceptable salt, thereof; or stereoisomers or tautomersthereof.
 5. A compound according to formula III;

wherein R¹ is H, or substituted or unsubstituted alkyl; R² is H, loweralkyl, lower cycloalkyl and lower alkyl-lower cycloalkyl, optionallysubstituted with one or more of F and Cl; R⁸ is

L is selected from a single bond, alkylene, —O—, —N(R^(8e))—, —CO—,—CO₂—, —SO—, —SO₂—, —CON(R^(8e))—, —SO₂N(R^(8e))—, —N(R^(8e))CO—,—N(R^(8e))SO₂—, —N(R^(8e))CON(R^(8e))—, —N(R^(8e))SO₂ N(R^(8e))—;—O(CR^(8e) ₂)_(m1)—, or —CON(R^(8e))(CR^(8e) ₂)_(m1)—; each R^(8a) isindependently selected from hydrogen, substituted or unsubstitutedalkyl, alkoxy, cyano, and halo; R^(8d) is selected from substituted orunsubstituted heterocycloalkyl; R^(8e) is selected from H, substitutedor unsubstituted alkyl and substituted or unsubstituted cycloalkyl; thesubscript m1 is 1, 2, 3, or 4; the subscript n is 1, 2, 3, or 4; andwherein R⁹ is

 or a pharmaceutically acceptable salt, thereof; or stereoisomers ortautomers thereof.
 6. A compound according to formula III:

wherein R¹ is H, or substituted or unsubstituted alkyl; R² is H, loweralkyl, lower cycloalkyl and lower alkyl-lower cycloalkyl, optionallysubstituted with one or more of F and Cl; R⁸ is

L is selected from a single bond, alkylene, —O—, —N(R^(8e))—,—CO—,—CO₂—, —SO—, —SO₂—, —CON(R^(8e))—, —SO₂N(R^(8e))—,—N(R^(8e))CO—,—N(R^(8e))SO₂—, —N(R^(8e))CO N(R^(8e))—, —N(R^(8e))SO₂ N(R^(8e))—;—O(CR^(8e) ₂)_(m1)—, or —CON(R^(8e))(CR^(8e) ₂)_(m1)—; each R^(8a)independently selected from hydrogen, substituted or unsubstitutedalkyl, alkoxy, cyano, and halo; R^(8d) is selected from substituted orunsubstituted heterocycloalkyl; R^(8e) is selected from H, substitutedor unsubstituted alkyl and substituted or unsubstituted cycloalkyl; thesubscript m1 is 1, 2, 3, or 4; the subscript n is 1, 2, 3, or 4; andwherein R⁹ is

or a pharmaceutically acceptable salt, thereof; or stereoisomers ortautomers thereof.
 7. A compound according to formula III:

wherein R¹ is H, or substituted or unsubstituted alkyl; R² is H, loweralkyl, lower cycloalkyl and lower alkyl-lower cycloalkyl, optionallysubstituted with one or more of F and Cl; R⁸ is

L is selected from a single bond, alkylene, —O—, —N(R^(8e))—,—CO—,—CO₂—,—SO—, —SO₂ —, —CON(R^(8e))—, —SO₂N(R^(8e))—, —N(R^(8e))CO—,—N(R^(8e))SO₂—, —N(R^(8e))CO N(R^(8e))—, —N(R^(8e))SO₂ N(R^(8e))—;—O(CR^(8e) ₂)_(m1)—, or —CON(R^(8e))(CR^(8e) ₂)_(m1)—; each R^(8a) isindependently selected from hydrogen, substituted or unsubstitutedalkyl, alkoxy, cyano, and halo; R^(8d) is selected from substituted orunsubstituted heterocycloalkyl; R^(8e)is selected from H, substituted orunsubstituted alkyl and substituted or unsubstituted cycloalkyl; thesubscript m1 is 1, 2, 3, or 4; the subscript n is 1, 2, 3, or 4; andwherein R⁹ is

 and wherein the subscript m is selected from 1-4 and each R^(9d) isindependently H, substituted or unsubstituted alkyl or halo; or apharmaceutically acceptable salt, thereof; or stereoisomers or tautomersthereof.
 8. A compound according to formula III:

wherein R¹ is H, or substituted or unsubstituted alkyl; R² is H, loweralkyl, lower cycloalkyl and lower alkyl-lower cycloalkyl, optionallysubstituted with one or more of F and Cl; R⁸ is

L is selected from a single bond, alkylene, —O—, —N(R^(8e))—, —CO—,—CO₂—, —SO—, —SO₂—, —CON(R^(8e))—, —SO₂N(R^(8e))—, —N(R^(8e))CO—,—N(R^(8e))SO₂—, —N(R^(8e))CO N(R^(8e))—, —N(R^(8e))SO₂ N(R^(8e))—;—O(CR^(8e) ₂)_(m1)—, or —CON(R^(8e))(CR^(8e) ₂)_(m1)—; each R^(8a) isindependently selected from hydrogen, substituted or unsubstitutedalkyl, alkoxy, cyano, and halo; R^(8d) is selected from substituted orunsubstituted heterocycloalkyl; R^(8e) is selected from H, substitutedor unsubstituted alkyl and substituted or unsubstituted cycloalkyl; thesubscript m1 is 1, 2, 3, or 4; the subscript n is 1, 2, 3, or 4; whereinR⁹ is

and wherein the subscript m is selected from 1-4and each R^(9d) isindependently H, substituted or unsubstituted alkyl or halo; or apharmaceutically acceptable salt, thereof; or stereoisomers or tautomersthereof.
 9. A compound according to formula III:

wherein R¹ is H, or substituted or unsubstituted alkyl; R² is H, loweralkyl, lower cycloalkyl and lower alkyl-lower cycloalkyl, optionallysubstituted with one or more of F and Cl; R⁸ is

L is selected from a single bond, alkylene, —O—, —N(R^(8e))—,—CO—,—CO₂—, —SO—, —SO₂—, —CON(R^(8e))—, —SO₂N(R^(8e))—, —N(R^(8e))CO—,—N(R^(8e))SO₂—, —N(R^(8e))CO N(R^(8e))—, —N(R^(8e))SO₂ N(R^(8e))—;—O(CR^(8e) ₂)_(m1)—, or —CON(R^(8e))(CR^(8e) ₂)_(m1)—; each R^(8a) isindependently selected from hydrogen, substituted or unsubstitutedalkyl, alkoxy, cyano, and halo; R^(8d) is selected from substituted orunsubstituted heterocycloalkyl; r^(8e) is selected from H, substitutedor unsubstituted alkyl and substituted or unsubstituted cycloalkyl; thesubscript m1 is 1, 2, 3, or 4; the subscript n is 1, 2, 3, or 4; whereinR⁹ is

and wherein the subscript m is selected from 1-3 and each R^(9d) isindependently H, substituted or unsubstituted alkyl or halo; or apharmaceutically acceptable salt, thereof; or stereoisomers or tautomersthereof.
 10. A compound of formula VIa, VIb, VId, or VIe:

wherein R^(8b) is hydrogen, substituted or unsubstituted alkyl orsubstituted or unsubstituted cycloalkyl.
 11. A compound of formula VIIa,VIIb, VIId, or VIIe:

wherein R^(8b) is hydrogen, substituted or unsubstituted alkyl orsubstituted or unsubstituted cycloalkyl.
 12. A compound of formulaVIIIa, VIIIb, VIIId, or VIIIe:

wherein R^(8b) is hydrogen, substituted or unsubstituted alkyl orsubstituted or unsubstituted cycloalkyl.
 13. A compound of formula IXa,IXb, IXd, or IXe:

wherein R^(8b) is hydrogen, substituted or unsubstituted alkyl orsubstituted or unsubstituted cycloalkyl.
 14. A compound of formula Xa,Xb, Xd, or Xe:

wherein R^(8b) is hydrogen, substituted or unsubstituted alkyl orsubstituted or unsubstituted cycloalkyl.
 15. A compound of formula XIa,XIb, XId, or XIe:

wherein R^(8b) is hydrogen, substituted or unsubstituted alkyl orsubstituted or unsubstituted cycloalkyl; and R^(9e) is hydrogen, Me, orCN.
 16. A compound according to any one of claims 10-15, wherein R^(8b)is H.
 17. A compound according to any one of claims 10-15, whereinR^(8b) is cycloalkyl.
 18. A compound according to any one of claims10-15, wherein R^(8b) is cyclopropyl.
 19. A compound according to anyone of claims 10-15, wherein R^(8b) is substituted or unsubstitutedalkyl.
 20. A compound according to any one of claims 10-15, whereinR^(8b) is Me, Et, Pr, i-Pr, t-Bu, i-Bu, CF₃, CH₂CF₃, CH₂CONH₂, orcyclopropylmethyl.
 21. A compound of formula XIIa, XIIb, XIIc or XIId:


22. A compound of formula XIIIa, XIIIb, XIIIc or XIIId:


23. A compound of formula XVa, or XVb:

wherein L is a single bond, —CO—,or —O—CH₂—CH₂—; the ring P is

and R^(8b) is H, Me, i-Pr, t-Bu, CH₂CONH₂, cyclopropylmethyl, or CH₂CF₃.24. A compound according to claim 23, wherein L is a bond and the ring Pis


25. A compound selected from(4-Morpholin-4-yl-phenyl)-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine;4-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-benzamide;[4-(4-Methylpiperazin-1-yl)-phenyl]-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine;4-{8-[4-(4-Methyl-piperazin- 1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide;Morpholin-4-yl-{4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-phenyl}-methanone;N-(2-Morpholin-4-yl-ethyl)-4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide;4-{8-[4-(2-Morpholin-4-yl-ethylcarbamoyl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzoicacid;4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-N-pyrrolidin-3-yl-benzamide;N-(1-Methyl-pyrrolidin-3-yl)-4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide;[4-(4-Methyl-piperazine-1-sulfonyl)-phenyl]-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine;4-{8-[4-(4-Methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-1H-pyridin-2-one;4-{8-[4-(4-Methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-thiophene-2-carboxylicacid amide;[4-(4-Methyl-piperazin-1-yl)-phenyl]-[5-(5-methyl-1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine;3-Methyl-4-{8-[4-(4-Methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide;2-Fluoro-4-{8-[4-(4-methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide;3-Fluoro-4-{8-[4-(4-methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide;2-Chloro-4-{8-[4-(4-methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide;5-{8-[4-(4-Methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-thiophene-2-carboxylicacid amide;4-(8-(3-chloro-4-(4-methylpiperazin-1-yl)phenylamino)imidazo[1,2-a]pyrazin-5-yl)benzamide;4-(8-(4-((2R,5S)-2,4,5-trimethylpiperazin-1-yl)phenylamino)imidazo[1,2-a]pyrazin-5-yl)benzamide;5-(8-(4-(4-methylpiperazin-1-yl)phenylamino)imidazo[1,2-a]pyrazin-5-yl)furan-2-carboxamide;2,6-difluoro-4-(8-(4-(4-methylpiperazin-1-yl)phenylamino)imidazo[1,2-a]pyrazin-5-yl)benzamide;4-(8(4-(4-isopropylpiperazin-1-yl)phenylamino)imidazo[1,2-a]pyrazin-5-yl)benzamide;4-(8-(6-(4-methylpiperazin-1-yl)pyridin-3-ylamino)imidazo[1,2-a]pyrazin-5-yl)benzamide;5-(8-(4-(4-methylpiperazin- 1-yl)phenylamino)imidazo[1,2-a]pyrazin-5-yl)thiophene-3-carboxamide;5-(8-(4-(4-methylpiperazin-1-yl)phenylamino)imidazo[1,2-a]pyrazin-5-yl)furan-3-carboxamide;2-fluoro-4-(8-(4-(2-morpholinoethoxy)phenylamino)imidazo[1,2-a]pyrazin-5-yl)benzamide;N-(4-(2-morpholinoethoxy)phenyl)-5-(1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-amine;5-(8-(4-morpholinophenylamino)imidazo[1,2-a]pyrazin-5-yl)thiophene-2-carboxamide;4-(8-(4-((3S,5R)-3,4,5-trimethylpiperazin-1-yl)phenylamino)imidazo[1,2-a]pyrazin-5-yl)benzamide;4-(8-(4-(4-(2-hydroxyethyl)piperazin-1-yl)phenylamino)imidazo[1,2-a]pyrazin-5-yl)benzamide;2-(8-(4-morpholinophenylamino)imidazo[1,2-a]pyrazin-5-yl)thiazole-5-carboxamide;2,6-Difluoro-4-[8-(4-morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-benzamide;4-(8-(3-fluoro-4-(4-methylpiperazin-1-yl)phenylamino)imidazo[1,2-a]pyrazin-5-yl)benzamide;4-(8-(3-morpholinophenylamino)imidazo[1,2-a]pyrazin-5-yl)benzamide;4-(8-(3-carbamoyl-4-morpholinophenylamino)imidazo[1,2-a]pyrazin-5-yl)-2,6-difluorobenzamide;5-(5-methyl-1H-pyrazol-4-yl)-N-(4-morpholinophenyl)imidazo[1,2-a]pyrazin-8-amine;4-(8-(4-morpholinophenylamino)imidazo[1,2-a]pyrazin-5-yl)thiophene-2-carboxamide;5-(8-(4-morpholinophenylamino)imidazo[1,2-a]pyrazin-5-yl)isoindolin-1-one;5-(8-(4-morpholinophenylamino)imidazo[1,2-a]pyrazin-5-yl)furan-3-carboxamide;2,6-difluoro-4-(8-(4-((2S,5R)-2,4,5-trimethylpiperazin-1-yl)phenylamino)imidazo[1,2-a]pyrazin-5-yl)benzamide;2-methyl-4-(8-(4-morpholinophenylamino)imidazo[1,2-a]pyrazin-5-yl)benzamide;4-(8-(4((2S,5R)-2,4,5-trimethylpiperazin-1-yl)phenylamino)imidazo[1,2-a]pyrazin-5-yl)thiophene-2-carboxamide;5-(8-(4((2S,5R)-2,4,5-trimethylpiperazin-1-yl)phenylamino)imidazo[1,2-a]pyrazin-5-yl)furan-3-carboxamide;5-[8-(4-Piperazin-1-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-furan-3-carboxylicacid amide;2,6-difluoro-4-(8-(4-(4-isopropylpiperazin-1-yl)phenylamino)imidazo[1,2-a]pyrazin-5-yl)benzamide;5-{8-[4-(4-Isopropyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-2,3-dihydro-isoindol-1-one;4-{8-[4-(4-Isopropyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-furan-2-carboxylicacid amide;4-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-furan-2-carboxylicacid amide;[6-(4-Isopropyl-piperazin-1-yl)-pyridin-3-yl]-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine;(5-Morpholin-4-yl-pyridin-2-yl)-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine;5-[8-(5-Morpholin-4-yl-pyridin-2-ylamino)-imidazo[1,2-a]pyrazin-5-yl]-2,3-dihydro-isoindol-1-one;4-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-2H-pyrazole-3-carbonitrile;5-{8-[6-(4-Isopropyl-piperazin-1-yl)-pyridin-3-ylamino]-imidazo[1,2-a]pyrazin-5-yl}-2,3-dihydro-isoindol-1-one;4-{8-[6-(4-Isopropyl-piperazin-1-yl)-pyridin-3-ylamino]-imidazo[1,2-a]pyrazin-5-yl}-furan-2-carboxylicacid amide; and pharmaceutically acceptable salts thereof
 26. A compoundselected from4-[3-Methyl-8-(4-morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-benzamide;[3-Methyl-5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-(4-morpholin-4-yl-phenyl)-amine;4-[3-Methyl-8-(4-morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-1H-pyridin-2-one;4-[3-Ethyl-8-(4-morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-benzamide;4-[3-Ethyl-8-(4-morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-1H-pyridin-2-one;[3-Ethyl-5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-(4-morpholin-4-yl-phenyl)-amine;[6-(4-Isopropyl-piperazin-1-yl)-pyridin-3-yl]-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine;5-[8-(6-Morpholin-4-yl-pyridin-3-ylamino)-imidazo[1,2-a]pyrazin-5-yl]-2,3-dihydro-isoindol-1-one;2,6-Difluoro-4-{8-[4-(2-morpholin-4-yl-ethoxy)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide;2-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-5,6-dihydro-furo[2,3-c]pyrrol-4-one;(3-Dimethylaminomethyl-4-morpholin-4-yl-phenyl)-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine;7-Fluoro-5-{8-[4-(4-isopropyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-2,3-dihydro-isoindol-1-one;2-{8-[4-(4-Isopropyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-5,6-dihydro-furo[2,3-c]pyrrol-4-one;5-{8-[4-(4-Isopropyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-3,3-dimethyl-2,3-dihydro-isoindol-1-one;2-{8-[4-(4-Isopropyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-4,5-dihydro-thieno[2,3-c]pyrrol-6-one;and pharmaceutically acceptable salts thereof.
 27. A compound selectedfrom(4-Morpholin-4-yl-phenyl)-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine;Morpholin-4-yl-{4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-phenyl}-methanone;N-(2-Morpholin-4-yl-ethyl)-4-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-ylamino]-benzamide;[4-(4-Methyl-piperazine-1-sulfonyl)-phenyl]-[5-(1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine;4-[8-(4-Morpholin-4-yl-phenylamino)-imidazo[1,2-a]pyrazin-5-yl]-1H-pyridin-2-one;4-{8-[4-(4-Methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-1H-pyridin-2-one;4-{8-[4-(4-Methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-thiophene-2-carboxylicacid amide;[4-(4-Methyl-piperazin-1-yl)-phenyl]-[5-(5-methyl-1H-pyrazol-4-yl)-imidazo[1,2-a]pyrazin-8-yl]-amine;2-Fluoro-4-{8-[4-(4-methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide;3-Fluoro-4-{8-[4-(4-methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-benzamide;5-{8-[4-(4-Methyl-piperazin-1-yl)-phenylamino]-imidazo[1,2-a]pyrazin-5-yl}-thiophene-2-carboxylicacid amide; and pharmaceutically acceptable salts thereof.
 28. Apharmaceutical composition comprising a pharmaceutically acceptablecarrier and a pharmaceutically effective amount of a compound of any oneof claims 25, 26 and
 27. 29. The pharmaceutical composition of claim 28,wherein the carrier is a parenteral carrier.
 30. The pharmaceuticalcomposition of claim 28, wherein the carrier is an oral carrier.
 31. Thepharmaceutical composition of claim 28, wherein the carrier is a topicalcarrier.
 32. A compound according to claim 4, wherein R⁸ is


33. A compound according to formula IVa, IVb, or IVc:

and wherein L is a single bond, —CO—,—O(CH₂)_(m1)—, —CON(H)(CH₂)_(m1)—,or —NHCO—; the subscript m1 is selected from 1-4; the ring P issubstituted or unsubstituted heterocycloalkyl; the subscript n, isselected from 1-4; each R^(8a) is independently selected from hydrogen,substituted or unsubstituted alkyl, alkoxy, cyano, and halo; and R⁹ is

and each of A¹, A² and A³ is independently selected from S, O, N,NR^(9a), and CR^(9a); each of R^(9a) is independently H or substitutedor unsubstituted alkyl; and R^(9b) is CONH₂, CONHMe, or CN; or apharmaceutically acceptable salt, thereof; or stereoisomers or tautomersthereof.
 34. A compound according to claim 4, wherein the compound isaccording to formula Va, Vb, Vd, or Ve:

and wherein R^(8b) is hydrogen, substituted or unsubstituted alkyl orsubstituted or unsubstituted cycloalkyl.
 35. A compound according toclaim 5, wherein R⁸ is


36. A compound according to formula IVa, IVb, or IVc:

and wherein L is a single bond, —CO—, —O(CH₂)_(m1)—, —CON(H)(CH₂)_(m1)—,or —NHCO—; the subscript m1 is selected from 1-4; the ring P issubstituted or unsubstituted heterocycloalkyl; the subscript n, isselected from 1-4; each R^(8a) independently selected from hydrogen,substituted or unsubstituted alkyl, alkoxy, cyano, and halo; and R⁹ is

 or a pharmaceutically acceptable salt, thereof; or stereoisomers ortautomers thereof.
 37. A compound according to claim 5, wherein thecompound is according to formula Va, Vb, Vd, or Ve:

and wherein R^(8b) is hydrogen, substituted or unsubstituted alkyl orsubstituted or unsubstituted cycloalkyl.
 38. A compound according toclaim 6, wherein R⁸ is


39. A compound according to formula IVa, IVb, or IVc:

and wherein L is a single bond, —CO—,—O(CH₂)_(m1)—, —CON(H)(CH₂)_(m1)—,or —NHCO—; the subscript m1 is selected from 1-4; the ring P issubstituted or unsubstituted heterocycloalkyl; the subscript n, isselected from 1-4; each R^(8a) is independently selected from hydrogen,substituted or unsubstituted alkyl, alkoxy, cyano, and halo; and R⁹ is

or a pharmaceutically acceptable salt, thereof; or stereoisomers ortautomers thereof.
 40. A compound according to claim 6, wherein thecompound is according to formula Va, Vb, Vd, or Ve:

and wherein R^(8b) is hydrogen, substituted or unsubstituted alkyl orsubstituted or unsubstituted cycloalkyl.
 41. A compound according toclaim 7, wherein R⁸ is


42. A compound according to formula IVa, IVb, or IVc:

and wherein L is a single bond, —CO—,—O(CH₂)_(m1)—, —CON(H)(CH₂)_(m1)—,or —NHCO—; the subscript m1 is selected from 1-4; the ring P issubstituted or unsubstituted heterocycloalkyl; the subscript n, isselected from 1-4; each R^(8a) is independently selected from hydrogen,substituted or unsubstituted alkyl, alkoxy, cyano, and halo; and R⁹ is

 and wherein the subscript m is selected from 1-4and each R^(9d) isindependently H, substituted or unsubstituted alkyl or halo; or apharmaceutically acceptable salt, thereof; or stereoisomers or tautomersthereof.
 43. A compound according to claim 7, wherein the compound isaccording to formula Va, Vb, Vd, or Ve:

and wherein R^(8b) is hydrogen, substituted or unsubstituted alkyl orsubstituted or unsubstituted cycloalkyl.
 44. A compound according toclaim 8, wherein R⁸ is


45. A compound according to formula IVa, IVb, or IVc:

and wherein L is a single bond, —CO—,—O(CH₂)_(m1)—, —CON(H)(CH₂)_(m1)—,or —NHCO—; the subscript m1 is selected from 1-4; the ring P issubstituted or unsubstituted heterocycloalkyl; the subscript n, isselected from 1-4; each R^(8a) is independently selected from hydrogen,substituted or unsubstituted alkyl, alkoxy, cyano, and halo; and R⁹ is

 and wherein the subscript m is selected from 1-4and each R^(9d) isindependently H, substituted or unsubstituted alkyl or halo; or apharmaceutically acceptable salt, thereof; or stereoisomers or tautomersthereof.
 46. A compound according to claim 8, wherein the compound isaccording to formula Va, Vb, Vd, or Ve:

and wherein R^(8b) is hydrogen, substituted or unsubstituted alkyl orsubstituted or unsubstituted cycloalkyl.
 47. A compound according toclaim 9, wherein R⁸ is


48. A compound according to formula IVa, IVb, or IVc:

and wherein L is a single bond, —CO—,—O(CH₂)_(m1)—, —CON(H)(CH₂)_(m1)—,or —NHCO—; the subscript m1 is selected from 1-4; the ring P issubstituted or unsubstituted heterocycloalkyl; the subscript n, isselected from 1-4; each R^(8a) is independently selected from hydrogen,substituted or unsubstituted alkyl, alkoxy, cyano, and halo; and R⁹ is

and wherein the subscript m is selected from 1-3 and each R^(9d) isindependently H, substituted or unsubstituted alkyl or halo; or apharmaceutically acceptable salt, thereof; or stereoisomers or tautomersthereof.
 49. A compound according to claim 9, wherein the compound isaccording to formula Va, Vb, Vd, or Ve:

and wherein R^(8b) is hydrogen, substituted or unsubstituted alkyl orsubstituted or unsubstituted cycloalkyl.
 50. A compound according to anyone of claims 7, 41, 42 and 43, wherein each R^(9d) is H.
 51. A compoundaccording to any one of claims 7, 41, 42 and 43, wherein m is 1 or 2 andeach R^(9d) is Me, Cl or F.
 52. A compound according to any one ofclaims 8, 44, 45 and 46, wherein each R^(9d) is H.
 53. A compoundaccording to any one of claims 9, 47, 48 and 49, wherein each R^(9d) isH.
 54. A compound according to any one of claims 8, 44, 45 and 46,wherein m is 1 or 2 and each R^(9d) is Me, Cl or F.
 55. A compoundaccording to any one of claims 9, 47, 48 and 49, wherein m is 1 or 2 andeach R^(9d) is Me, Cl or F.